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B Total counts of WT donor CD4+ T cells in mice from (A) (n=13 100% WT, 8 25% WT, 14 10% WT, 6 1%, 14 0% WT). Two representative experiments shown out of 3. Mean ± SEM. | [
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(C) The indicated plasmids of Myc-tagged wild-type NLRP3 or its mutants Y132F, Y164F, Y251F, Y570F and Y589F were respectively transfected into HEK293T cells along with GFP-tagged EphA2, and then cell lysates were immunoprecipitated with the anti-Myc antibody, followed by western blot analysis with the anti-p-Tyr antibody, anti-HA antibody and anti-Myc antibody. | [
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Human osteosarcoma U2OS cells were pre-treated with dactinomycin (DACT), bortezomib (BTZ), daunorubicin (DAUN), docetaxel (DOC), doxorubicin (DOXO), epirubicin (EPI), mitoxantrone (MTX), paclitaxel (PACL), vinblastine (VB) and vincristine (VC) at 0.5, 1 and 5 µM ; with cisplatin (CDDP) at 75, 150 and 300 µM, with oxaliplatin (OXA) at 250, 500 and 1000 µM and with crizotinib (CRIZ) at 10, 20 and 40 µM for 1.5 to 2.5 h Cells were treated for 2.5 h and before fixation and permeabilization. Then, cells were stained with a rabbit anti-fibrillarin antibody followed by a staining with an anti-rabbit AlexaFluor-647- or AlexaFluor-546-coupled secondary antibody as well as with a mouse anti-nucleolin antibody followed by a staining with an anti-mouse AlexaFluor-488-coupled secondary antibody. Then images were acquired and colocalization between both signals was assessed (C). The surface overlap coefficient (SOC) was calculated and ranked between the untreated control (Ctr) and the positive control (DACT) Representative images of DACT 1 µM, BTZ 1 µM, CDDP 150 µM, CRIZ 20 µM, DAUN 0.5 µM, DOC 1 µM, DOXO 1 µM, EPI 1 µM, MTX 1 µM, OXA 500 µM, PACL 1 µM, VB 1 µM, VC 1 µM are shown | [
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A. Top: Molecular architecture of tetravalent F4L5.13. Bottom: Schematic for activation of Wnt/ßcatenin signaling by F4L5.13. | [
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Bone marrow from wild-type (Atg16L1+/+) was used to reconstitute irradiated littermate control mice (B6 WT → control [●] ) or δWD mice (B6 WT → δWD [○] ). Bone marrow from δWD mice was used to reconstitute irradiated δWD mice (δWD → δWD [▲] ). After 12 weeks, mice (n = 5 per group) were challenged with IAV X31 (103 pfu). (D) Lungs taken at 5 d.p.i. were analysed for neutrophils (Ly6G), neutrophil extracellular traps (NET; anti-H3) and macrophages (Iba-1) . | [
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Time-lapse confocal imaging (at a rate of one image stack/min) was used to document dynamic behavior of ONL microglia in rd10 retinal explants at ages P21-24. Example in which the engulfment of photoreceptorsomata by microglial phagocytic cup was followed up by actual phagocytosis in which the engulfed cell (red *) is translocated intracellularly within the microglial cell toward the cell body. Phagocytosis of soma occurred simultaneously with "probing" of other somata by processes of the same microglial cell (yellow *). | [
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B. Illustration outlining APEX2-catalyzed biotin-phenol labeling. An Rbd2-Flag-APEX fusion protein oxidizes biotin-phenol to biotin-phenoxyl radical that covalently labels binding partners. | [
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E. Comparison of the position of Arp2 relative to Arp3 in the short pitch arrangement (grey Arp2), the inactive splayed arrangement (red Arp2), and the SPIN90 bound x-ray crystal structure (cyan). The short pitch arrangement of Arp2-Arp3 was modeled by overlaying subdomains 1 and 2 of Arp3 on the same subdomains in an actin subunit in the actin filament structure 3G37(Murakam ), and then superposing Arp2 from the 4JD2 structure onto the actin subunit in the short pitch position. The SPIN90-bound complex was then overlaid onto Arp3 subdomains 1 and 2 to determine the relative position of Arp2 in the SPIN90-bound complex. Blue line shows a potential trajectory of the center of mass of Arp2 when it moves from the splayed to the short pitch position | [
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A Bacterial pathogens invade leaf tissues through stomata. B Pathogens proliferate in the extracellular space or the apoplast. C, D Virulent pathogens spread over the infection site (C) and develop disease (D). E, F Avirulent pathogens are restricted to the infection site as a result of the construction of the CASPL- and lignin-deposited structure (E) and are eventually eliminated from the infection site of the leaf (F). | [
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(A) Experimental design. Yeast cells growing on glucose were switched to no carbon media for 30 min. Thereafter, 20 mM H2O2 was added and metabolome analyzed by LC‐MS. | [
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(D) CP20 and OVCAR4 stable cell lines were transfected as either empty vector (EV) or pLYS1-MICU1-Flag (MICU1) and relative MICU1 levels were evaluated using immunoblotting. | [
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C) Thermal shift stabilization of purified dimeric NHE9*-GFP in the presence of PIP2 (red) compared to PIP2-free (black). Data presented are normalized mean FSEC peak fluorescence as mean values ± data range of n = 2 technical repeats; the apparent Tm was calculated with a sigmoidal 4-parameter logistic regression function; the average ΔTm presented is calculated from n = 2 independent titrations. | [
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A. Protein sequences were extracted from UniProt accession number BP08_BPT4 for Enterobacteria phage T4 (gp8; Bacteriophage T4) and the Pseudomonas Genome Database (TssK1; reference strain PAO1). Conserved positions are shown in black and grey background. The secondary structural elements corresponding to the 3D structure of gp8 (PDB code 1PDM) are shown above the alignment. Regions with significant sequence conservation are indicated with a corresponding block number. | [
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Paraffin sections from Bouin-fixed wild type and SPPL2c-/- testis were stained with hematoxylin & eosin (H&E). Scale bars, 100 µm. | [
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C RNAlow neoblast size in intact animals and at 48 hpa in control or TOR KD animals (n ≥ 55). | [
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B. Representative images of growth cones of mouse DRG neurons transfected with WT or mutant constructs treated with 75 nM Sema6A-Fc and visualized with immunofluorescent staining to detect GFP (green) and phalloidin (red). Average intensity images for all different constructs are shown in grey color. Scale bar 20 µm. | [
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(G) Hel2(1-315) polyubiquitinates eS7A but not uS10. Western blotting after an in vitro ubiquitination assay using the Hel2-(1-315) mutant and purified ribosomes containing HA-tagged uS10. | [
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Volcano plot of differential transcription factor (TF) motif accessibility (activity) in GPR56high (turquoise) vs GPR56low (violet) samples and their corresponding adjusted p-values determined with diffTF. Highlighted are TFs whose RNA expression was also positively or negatively affected by GPR56 KD in the RNA-seq dataset. | [
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Simulation of the PFK rate with varying ADP/(ATP+ADP) ratios. Data information: The PFK flux was simulated as single (isolated) reaction with the following fixed metabolite concentrations relevant for the kinetic rate law of the PFK PEP: 0.27 µmol/gDW; F6P: 0.91 µmol/gDW; FBP: 9.74 µmol/gDW ; total concentration ATP+ADP: 2.67 µmol/gDW. | [
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Upper part: in resting cells (no stress), Hsp90 interacts with DYRK3 and regulates its stability and activity. Upon stress, DYRK3 dissociates from Hsp90 and partitions inside SGs, where it is protected from irreversible aggregation and degradation. During the recovery phase after stress, soluble DYRK3 associates with Hsp90 outside or at the boundary of SGs to be stabilized. Once stabilized by Hsp90, DYRK3 could autophosphorylate its activation loop to achieve full activation and trigger the disassembly of SGs, by phosphorylating yet unknown molecular targets. Active DYRK3 would then promote translation restoration. Lower part: upon treatment with GA or 17AAG, DYRK3 dissociates from Hsp90, becomes inactive and is either targeted to SC35 speckles or degraded by the 26S proteasomes. During stress, blocking the loading of DYRK3 onto Hsp90 with GA or 17AAG redirects DYRK3 to SGs or to 26S proteasomes. During the recovery phase after stress, inhibition of Hsp90 prevents the stabilization and activation of DYRK3. As a consequence, SGs do not disassemble and translation is not restored. The model only depicts the interplay between Hsp90 and DYRK3 in the regulation of SG dynamics, although other Hsp90-dependent/DYRK3-independent mechanisms may also occur. | [
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H. Immunofluorescence images against CB1 and the Purkinje cell marker calbindin in the cerebellum of age-matched control and ASMD-affected children. Graph shows mean ± SEM intensity associated to CB1 in the Purkinje cells expressed as percentage of the values obtained in the control child (16 and 15 replicates in control and ASMD, respectively). Scale bar, 10 μm. | [
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(c) Juxtanuclear MRs are found in Rab7- and LAMP2-positive lysosomes. U2OS cells stably expressing DsRed-Rab7 were fixed and stained for MKLP1 α-tubulin and DAPI (left panels); wild-type U2OS were stained for MKLP1 and LAMP2 (right panel). Scale bars are 10 μm. The insets are enlargements of the engulfed MRs. Scale bars are 1 μm. | [
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A Levels of 4EBP1 protein and its phosphorylation at Serine 65 in presence of DMSO, rapamycin (100 nM) or temsirolimus (100 nM). GAPDH is showed as loading control. | [
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E. Comparison across treatment cohorts of the relative change in baseline of Per2-driven bioluminescence when vehicle, Prok2 or VIP was applied within CT9-12. Only VIP is statistically shown to significantly raise Per2::Luciferase baseline levels (p=0.0007; vehicle vs. Prok2, ns. p=0.17; vehicle vs. VIP, **p=0.002, Prok2 vs. VIP, *p=0.02). F. The relative change in amplitude of Per2::Luciferase rhythm after treatment with Prok2 or VIP significantly decreases compared to vehicle application (p=0.002; vehicle vs. Prok2, *p=0.01, vehicle vs. VIP, **p=0.001). | [
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Phosphorylation of Ub by PINK1 has multiple consequences. Structurally, it alters the electrostatic potential of Ub, but also generates a new UbretraCT conformation with a retracted C‐terminus. The UbretraCT conformation is in slow exchange with the common Ub conformation. Functionally, Ub phosphorylation leads to a gain‐of‐function of Ub, as it becomes an allosteric activator of Parkin. We here show that phosphoUb also has loss‐of‐function effects, as it inhibits some assembly and disassembly systems. Additionally, it is possible that phosphoUb is recognized specifically by Ub receptors. | [
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D Oocytes expressing KCC4 were incubated with NAM for the indicated times, and KCC4 protein levels (upper panel), and activity (lower panel) were analyzed by SDS-PAGE/immunoblotting or Cl--dependent 86Rb+ uptake under hypotonic conditions. | [
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A Close-up view of Ca2+-free MICU1-MICU2 interfaces I (left and middle) and II (right). The atomic interactions at the interface I consist of mainly electrostatic (left) and weak hydrophobic (middle) interactions, while interface II mainly contains hydrophobic interactions (right) | [
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I: Relative mtDNA copy number in Detroit 551, WS5A and CP2A iPSCs by RT-qPCR analysis using ND1 and APP (n=5, technical replicates per clone for control; n=5, technical replicates per clone for WS5A and CP2A). Values are presented as Log2 of the ratio between the expression values of ND1 in relation to APP. | [
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D, multiscale network of cell sub-community and coEM eigengenes depicted with a force-directed layout and edge bundling. Gold nodes, cell sub-communities; gray nodes, coEM eigengenes; large diamonds, clinical variables; red edges, positive correlations; blue edges, negative correlations. Edges are filtered to Pearson correlations significant at P < 0.001, and thickness corresponds to the square of the correlations. A cluster of sub-communities and coEMs associated with convalescence (dashed gray box) and a corresponding cluster of sub-communities and coEMs associated with acute infection (solid black box) surround the "convalescent" node. A positive correlation between convalescent anti-CHIKV antibody (Ab) titer and CD14+CD16+ monocyte sub-community 1 frequency (shown previously in Appendix Figs S3A and S3B) is also visible (asterisk and arrow). | [
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H. Lysates from HeLa cells were subjected to ChIP assay. ChIP products were amplified by qPCR reaction with the indicated pairs of primers. Data shown are mean ± SD (n = 3; *P < 0.05, two-tailed t-test). | [
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G Quantification of FLAG and DAPI fluorescence intensity along a line centered at the nuclear envelope. Eight OHC were measured. | [
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Micrograph and LC3B antibody labelling of adipocytes neighboring the nerve cells of Porcellio d. dilatatus.These adipocytes are localised around the nerve cells of the CNS and constitutes the sheathing of this organ. (A) At 30 days post-injection, autolysosomes, autophagosomes but also bacteria were observed: autophagic process was already higher compared to the control (Ad: adipocyte, Ap: Autophagosomes (Autophagic hallmark), Al: Autolysosomes, Wo: Wolbachia, Arrow head: magnified area). (B) At 60 days post-injection, the adipocytes were severely damaged by the accumulation of autophagic vesicles (Ad: adipocyte, Ap: Autophagosomes, Al: Autolysosomes, Wo: Wolbachia, Arrow head: magnified area). (C) With the antibody labelling anti-LC3B, the protein LC3B implicated in autophagic processes was detected in adipocytes of P. d. dilatatus injected with wVulC at 45 days post-injection. The highly labeled spherical structures would reflect the incorporation of the LC3B protein in the phagophores. (D) With adipocytes of P. d. dilatatus injected by wDil but also in other tested situations, only really few of these structures were detected by epiflorescent microscopy. | [
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A. Schematic of the differentiation protocol timeline. Maintenance Medium (MM) = iPSC medium (StemFlex with 1x penicillin/streptomycin), NCC differentiation medium = DMEM-F12, 10% fetal bovine serum, 1 mM sodium pyruvate, 1 mM penicillin/streptomycin, 1 mM non-essential amino acids, 110 µM 2-mercaptoethanol, 10 ng/mL epidermal growth factor. | [
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Differential expression analysis for ACE2 expression changes in treated vs control samples was performed with the level 3 CMAP data of 24 hours response values using limma (Ritchie et al, 2015; Methods). Volcano plots showing the log fold-change (X-axis) and uncorrected negative log10P value (Y-axis) of ACE2 expression changes are given for (A) 48 antihypertensive drugs The enrichment of positive/negative ACE2 expression regulators in different drug classes based on their mechanism of action (MOA) was tested with the GSEA method as implemented in the R package fgsea (Segushichev, 2016; Methods), and the enrichment significance (negative log10 P values) is shown in bar plots in (B) , for the analysis on the 48 antihypertensive drugs CCBs, calcium channel blockers; ARBs, angiotensin II type-I receptor blockers; ACEIs, angiotensin-converting enzyme inhibitors | [
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(D) Western blotting analyzed Il-17rb in shLacZ or shTgfbr1 4T1 cells treated with recombinant TGF-β1 for 5 days. | [
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UMAP plot revealing the cell type annotations of three samples. B) The cellular composition of three healthy kidney samples. | [
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E Western blot analyses showing the extent of HIF1α stabilization and steady state level of VHL protein with and without the treatment with FG4592 (100 μM) in RKO cells. GAPDH is showed as loading control. | [
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J Time course of expression of WT-CD40LG mRNA or CO-CD40LG mRNA, measured as fold change (FC) on IPO8 housekeeping gene and normalized to T0. Sorted edited (+) and sorted unedited (-) CD4+ T cells were analyzed before and 3, 6, 12 hr after Actinomycin D treatment (n=1 for each group). | [
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(H-J) Blood ketone body levels 6h after normoxia and hypoxia in female C57BL/6J and ADX mice (H), in female C57BL/6J mice injected with 5 mg RU486 or vehicle (DMSO) (I), and in GRdim/dim mice and their wild-type littermates (J). N=5-9 per group. Data information: All bars represent mean ± SEM. P-values were calculated using two-way ANOVA followed by post-hoc Šídák's multiple comparisons test to correct for multiple testing during the pairwise multiple comparisons, except if otherwise stated. ****P<0.0001, ***P<0.001, **P<0.01, *P≤0.05. | [
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BsAb were incubated at the indicated concentrations with PBMC of healthy donors together with 22Rv1low-cells expressing high and low amounts of PSMA, respectively. After three days, cytokine release were determined by flow cytometry as described in the methods section. Mean values and standard deviations of triplicate measurements are indicated. | [
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(B, C) Wild‐type and HDAC6 KO MEFs were transfected with mCherry‐GFP‐LC3, followed by treatment with LatA (100 nM) or nocodazole (250 nM) for 6 h and analysed as described in Figure 3A. | [
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(C) Representative traces obtained with ADP•BeFx. The upper FRET trace was calculated from the middle traces obtained by exciting the donor fluorophore and measuring both donor (green) and acceptor (red) fluorescence. The lowest trace was obtained by exciting the acceptor fluorophore directly. The arrow indicates a bleaching event. (D) Distribution of FRET values determined from 97 traces as in (C) fit with a Gaussian model (black curve). | [
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Percentage of early apoptotic EMPs and Early E in wild type and Sl/Sl E11.5 FL, determined by flow cytometric analysis of AnnexinV staining among 7-AAD- Ter119- CD41+ Kit+ CD16/32+ EMPs and 7-AAD- Ter119- Kit+ CD41- CD71+ CD44+ Early E. Data are the mean (±SD) of 9 wild type and 7 Sl/Sl biological replicates analyzed over 3 independent experiments, with each replicate consisting of 1-3 FLs of identical genotypes. A total number of 13 wild type and 9 Sl/Sl embryos were analyzed. Tail somite range: 11-17 for both genotypes. | [
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Representative WB analysis of pSer+pThr and GADPH (d) expression in the TA and VM muscles of C57Bl6/J and mdx mice (n=10 mice per group). Densitometry analysis of WB data are expressed as the pSer+pThr/CD20 ratio in arbitrary units in the lower panels One-way ANOVA. ****p<0.0001. | [
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Log2 fold change (LFC) in TAUtg (x-axis) and APPtg mice (y-axis) after differential expression analysis. Upregulated genes are on the right part (TAUtg mice) or upper part (APPtg mice) of the graph; downregulated genes are on the left part (TAUtg) or lower part (APPtg) of the graph. Colored dots represent significantly differentially expressed genes (Benjamini-Yuketieli-adjusted p-values (Padj) <0.05) for APPtg (green dots), TAUtg (yellow dots) or for both (red dots). Spearman correlation assesses the correlation between APPtg and TAUtg mice when ranking genes that are significantly differentially expressed in either APPtg or TAUtg mice from most up- to most down-regulated on a combined score of LFC and padj (i.e. signed log10(p-value), where the sign is determined by the LFC). A) Genes differentially expressed as a function of age, i.e. 4M vs 10M, independently of genotype. Thus, genes with a positive LFC are more highly expressed in 10M mice over 4M mice. | [
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The percentage that Lipin, Ctdnep1, and Nep1r1 RNAi normalize the abundance of membrane GL and GPL classes in the dTorKO fat body. 0% "normal" reflects lipid abundance in dTorKO expressing Luciferase RNAi, while 100% "normal" reflects lipid abundance in w- expressing Luciferase RNAi. Percentage normalization was separately calculated for PI, PA, DAG, PC, PE and PS (for dTorKO expressing Lipin, Ctdnep1, and Nep1r1 RNAi). Points show the mean of all lipid class normalization per sample, bars show mean ± SEM for the group. | [
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A Genomic attributes of RNA associated with renatured NF-H, Tau, or Aβ. Inset shows which fractions of the peaks that derive from exons or introns have their origin in coding or non-coding transcripts, respectively. | [
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Analytical SEC chromatograms and corresponding Coomassie stained SDS-PAGE gels of Cse4n (light blue), Mif2-wt (red), combination (green). Note that the same Cse4n (upper panel) and Mif2 (middle panel) elution profiles and SDS-PAGEs are also displayed as reference in B and C (Mif2) or C (Cse4n) to improve clarity. | [
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Co-immunoprecipitation of exogenous FLAG-USP28 C171A and FLAG-ΔNp63 in HEK293 cells. ΔNp63 was precipitated and blotted against FLAG-USP28 or ΔNp63. The input corresponds to 10% of the total protein amount used for the IP (ACTIN as loading control). | [
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(A) (Ai) Western blot analysis was used to confirm UHMK1 overexpression and ATF4 knockdown in BGC823 cells. (Aii) Silencing ATF4 significantly decreased the levels of the indicated metabolites in BGC823 cells overexpressing UHMK1(three biological replicates). (Aiii) Silencing ATF4 significantly decreased the levels of RNA and DNA containing U-14C-glycine in BGC823 cells overexpressing UHMK1(three biological replicates). (B) LC-MS/MS was used to examine the metabolites in SGC7901 cells with or without UHMK1 knockdown or ATF4 reintroduction. The data are shown in the heatmap. (C) Silencing ATF4 significantly decreased the levels of the indicated genes in BGC823 cells overexpressing UHMK1. Data were presented as mean ± standard deviation from three replicates. Unpaired, two-tailed statistical significance was assessed by Student's t‐test. (D) (Di) qRT-PCR was used to measure the expression of ATF4 in BGC823 cells overexpressing UHMK1. (Dii) UHMK1 overexpression enhanced the nuclear translocation of ATF4 in BGC823 cells. (Diii) ChIP assays on the ATIC and PPAT promoters were performed in BGC823 cells with the indicated antibody. Data were presented as mean ± standard deviation from three replicates. Unpaired, two-tailed statistical significance was assessed by Student's t‐test. Data information: *P< 0.05, **P< 0.01, ***P< 0.001, # marked no significance. | [
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a, iCAF T-cell dysfunction gene signature highlighting genes significantly associated with CTL dysfunction in two out of three independent patient cohorts (METABRIC, GSE21653 and GSE58812). | [
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C Dual immunohistochemistry staining of human breast tumor with p-EDC3 Ab (Fredriksson et al.) and total EDC3 (brown). Scale bar: 50µm. D Dual immunohistochemistry staining of human normal resected breast tissue sections with p-EDC3 Ab (Fredriksson et al.) and total EDC3 (brown). Scale bar: 50µm. | [
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(E) Quantification of one set of representative immunoblots (in figure D) and its graphical representation showing dynamic of Smad2 and JNK activation during TGF-β-induced EMT in NMuMG. | [
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D-F RT-qPCR analysis of interferon-related genes (D), western blot analysis of TERT, TBK1, pTBK1, IRF3, pIRF3, and GAPDH levels (E), and ELISA of IFNβ and CXCL10 in culture supernatant (F) in U2OS and HeLa cells ectopically expressing TERT-WT or TERT-K626A. | [
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B Bar graph showing the relative abundance of phyla from each mouse cecum, where each bar is a separate mouse. | [
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C Local resolution map of the refined Apo and AMP-PNP particle, as calculated with Blocres (Heymann & Belnap, 2007). | [
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(I-J) Relative RNA abundances for Siglec-8 in (H) hippocampus and (I) posterior cingulate cortex post-mortem patient tissue. Linear regression R2 values are shown on plots. | [
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P Kaplan-Meier curve comparing relapse-free survival of primary tumors with high and low values of UBE4B. P value was calculated using log-rank (Mantel-Cox) test. | [
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(K-M) Immunofluorescence images (K) and quantification of the percentage of ciliated cells (L, n = 3 mice) and ciliary length (M, n = 100 cilia from three mice) in mouse kidneys stained with the antibody against acetylated α-tubulin and DAPI. To quantify the percentage of ciliated cells (L), >200 cells from 12 images were analyzed for each mouse. Scale bar, 3 µm. | [
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PCR array analyses were performed on LCM captured P7 CLE of Prom1 and Glis2 KO vs. WT mice (n = 3). Arrows indicated genes with expression changed for more than 2 folder increase or decrease. | [
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(C) Quantification of engulfed mCherry+ materials in microglia. N = 5 for biological replicates. | [
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(A) Schematic showing experimental setup to measure rates endocytosis in BV-2 cells. Control (MIP empty vector) cells were plated together with cells stably expressing different Siglec constructs. Cells are allowed to adapt to the same media environment for 24 hours, then fluorophores are added to media. Cells with endocytosis substrates (Aβ, Dextran) were assayed after 3 hours, while phagocytosis substrates (FluoSpheres) were assayed after 24 hours. Populations are separated and relative uptake is measured using flow cytometry. | [
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(H) U2OS PIF1-KO cells expressing PIF1 WT or E307Q, L319P mutants were labeled with CldU for 30 minutes, followed by 2 mM HU treatment for 2 hr and IdU labelling for 30 minutes, and processed for DNA fiber analysis. The percentage of replication restarted DNA fibers is calculated. Data information: Error bars represent the standard deviation (SD) of at least three independent experiments. Significance of the differences was assayed by Two-tailed non-paired parameters were applied in Student's t-test. The P value is indicated as *P < 0.05, **P < 0.01, ***P < 0.001 and n.s. (not significant) P>0.05. | [
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(G) Immunoblots for Parkin, PINK1, phosphorylated p-62 (p-p62), HSP60, LC3, p62/SQSTM, and Atg5 using the mitochondrial fraction and whole cell lysates of Huh7 cells. HSP60 and β-actin were used as loading controls for mitochondria and whole cell lysates, respectively. | [
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Confocal fluorescence images of the differentiation and division zones of Arabidopsis root tips expressing pSUC2::HST:GFP, pSUC2::GFP or pSUC2::tmGFP9. Scale bars: 100 µm. Confocal fluorescence images of the differentiation and division zones of pSUC2::GFPhst-1 or pSUC2::tmGFP9hst-1 root tips. Scale bars: 100 µm. | [
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E, Experimental design to measure GSC frequency in tumors treated as in C (JNJ was administered for 10 days). Explanted tumors were dissociated and viable cells were tested by in vitro sphere forming LDA or in vivo single-passage (p1) LDA. | [
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A UMAP plots focusing on DN3a, DN3b γδ+ and DN4 γδ+ cells from Dataset 1. Clustering was performed using a 1.5 resolution. The UMAP plot is colored according to clusters and the main characteristics of the defined clusters specified in the right margin. | [
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A Representative microCT images of trabecular bone from the femoral metaphysis of male Sp7-Cre;Usp34f/f and littermate control mice. Scale bar, 500 μm B, C Von Kossa stainin | [
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D. Transmission electron microscopy of a collagen matrix section. Scale bar = 1 µm. Positions of magnified areas indicated by corresponding colored squares. | [
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A) Confocal image of wing disc expressing the human tetraspanin CD63-GFP expressed in the Hh receiving cells colocalizing with Ptc (Bac.Ptc-cherry) in vesicle-like structures (yellow arrows) along basal cytonemes. | [
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E. Representative images of mousehippocampal neurons transfected at 4 DIV with either GFP-expressing control vector or Lrig1shRNA-GFP plasmid. After transfection, neurons were cultured in the absence or in the presence of BDNF (25 ng/ml) for 48 h (7 DIV). Arrows indicate branching points along the principal dendrite. Scale bar, 10 m. | [
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(D) Meiotic mRNAs accumulate in conditional mmi1Δ cells. Total RNA samples from mei4‐P527, red1Δmei4‐P527, and mmi1Δmei4‐P527 cells were subjected to RT-PCR analyses. RT-PCR of mei4+, mcp5+, and SPBPB2B2.03c indicates that red1Δ or mmi1Δ in a mei4‐P527 background results in increased levels of these meiotic transcripts. Note that mmi1+ was deleted in the mei4‐P527 mutant strain because mei4 deficiency suppresses the severe growth defect caused by mmi1Δ. | [
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D Respiration of Subject 3 and Subject 1 derived fibroblasts (black and grey, respectively) compared to control fibroblasts (white) using high-resolution respirometry. ROX-corrected analysis of basal respiration, Oligomycin inhibited LEAK respiration and maximum uncoupled respiration (ETS). Both subject fibroblasts exhibited a severe mitochondrial respiration defect as measured by oxygen consumption. Data are mean ± SD from at least 3 experiments and analysed by Student's t-test. **p<0.01, ***p<0.001. | [
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(D) Time-lapse observation of the establishment of spindle bipolarity upon NuMA depletion. HCT116 TetOsTIR1 NuMA-mAID-mClover-FLAG cells with 2-centrosomes were treated with 1 μg/ml Dox and 500 μM IAA and observed with a 40× objective. Red and green represent SiR-tubulin and NuMA, respectively. Z-projections of 14 sections, every 2 μm. Scale bar, 10 μm. Time zero corresponds to NEBD. (E) The time required for the initial establishment of spindle bipolarity in (D). Line and error bars represent the mean and SD. (N≧60 cells from two independent experiments). The Mann-Whitney U test (two-tailed) was used to obtain a P value. | [
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A HeLa cells were treated with control (CTRL), SEPT7, or Drp1 siRNA for 72 h. Whole cell lysate of siRNA-treated cells were immunoblotted for GAPDH, SEPT7, or Drp1 to show the efficiency of siRNA depletion. GAPDH was used as loading control. siRNA-treated cells were labelled with MitoTracker Red CMXRos, and fixed for confocal microscopy. The scale bar represents 5 μm. | [
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An orthotopic lung tumor model was generated using A549 human lung cancer cells and treated as in (C). Figure displays H&amp;E staining of lung sections from radiation-treated (upper panel) and RK-33- and radiation-treated mice (lower panel). Scale bar is 2 mm.Quantification of tumor burden (as tumor surface divided by total lung surface) in orthotopic A549 lung cancer mouse model, as shown in (F). Significance was assessed by two-sided, unpaired t-test. Error bars represent SD. | [
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(E) Coimmunoprecipitation of Pex3‐177 with Atg36. IP was performed in the background strain of C13-Atg36-PtA using a plasmid expressing Pex3‐177-GFP under control of its endogenous promoter. Cells grown for 24 h to post‐log phase in glucose medium. IgGSepharose beads were used to immobilise Atg36-PtA from spheroplastyeast lysates. SDS-PAGE gels were probed with anti‐GFP and PAP. Yeast lysates represent 5% of the lysate added to the beads and analysed by immunoblotting. TL, total lysate; IP, immunoprecipitate. Comparison to Pex3-GFP IP (left panel). | [
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IBA-1 (green staining, RPE far-red autofluorescence) double-labeling of the GA lesion shown in (H). Representative images from five donor eyes with large drusen, five donor eyes with GA, and three healthy donors between 70 and 89 years old. | [
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E) Co-immunostaining of endoplasmic reticulum (ER) marker (KDEL, in red), thyroglobulin (Tg in green), and both merged (from left to right) in adult thyroid tissue. Note the thyrocyte disorganisation in Tubb1-/- mice | [
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K, L Western blot analysis of CD38 protein expression in heart (K) and diaphragm (L) of WT (n=5, n=6, respectively) and mdx (n=5) mice. Vinculin is used as housekeeping protein control and the dot plots show the ratio of CD38/vinculin. Data information: Each dot of the graphs represents a value/mouse. After normality and variance comparison tests, significance was assessed using: unpaired Student's t-test; Values are expressed as means ± SEM. Significance: *p<0.05, **p<0.01, ***p<0.001. | [
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A. Osmotic stress induces YAP nuclear translocation. HEK293A cells were serum starved for 1 hr followed by 0.2 M sorbitol treatment for 1 hr. YAP/TAZ (red, upper panels) or YAP (red, lower panels) were stained with two specific antibodies. DAPI (blue) was used to visualize cell nuclei. Scale bar: 20 μm. Quantification of more nuclear (N > C) or more cytosolic (N < C) YAP signal is determined with randomly chosen fields. | [
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Schematic representation of initial events of IAHC formation. (i) A single cell protrudes into the lumen of the aorta and starts expressing CD41. (ii) The CD41:YFP+ bulging cell undergoes mitosis (iii), forming the core of the IAHC. | [
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P. Percentage of normal, abnormal and missing rhabdomeres for the genotypes mentioned above; n=180 ommatidia from 3 different fly retinas for each. Data were quantified for significance using Student's t test. **p<0.01. | [
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J-N. Images show a wild-type plant, CDKA;1-DE and the double mutant cdkb1;1 cdkb1;2 (from left to right) on the indicated day after germination and the indicated drug treatment. | [
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A, B RT-qPCR analysis of transcript levels of TERRA (A) and tlh1+ (B) in WT strain and mutants as indicated (double mutants with ccq1Δ are indicated by blue dot). Data information: In (A, B), data is represented as mean ± SEM from 3 independent experiments and shown relative to WT leve | [
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C, D: U2OS cells were transfected with EGFP-tagged p14ARF alone or in combination with Myc-tagged wild type PRMT1-containing plasmids. p14ARF-EGFP (green), DAPI (blue, nuclei/DNA) and PRMT1 (red, using α-Myc antibody) were visualized by fluorescence microscopy, for which representative results are shown in (C), with the lower images displaying a magnification as indicated in the upper images by the rectangles. Scale bars: 10 μm. The subcellular distribution of p14ARF-EGFP (exclusively nucleolar or not-exclusively nucleolar but predominantly nucleo-/cytoplasmic) was quantified in p14ARF-positive and p14ARF/Myc-PRMT1-double positive cells by cell counting (percentage of cells) for three independent experiments in (D) (mean ± SD, ***: p-value ≤ 0.001 using Welch´s t-test). | [
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(a-c) Hematoxylin and eosin staining (a) of the SVZ and dentate gyrus (DG) of P28mice treated by NAC or vehicle (Veh) control. Dotted lines indicate boundaries for the SVZ and DG. Arrows mark cells in the SVZ. | [
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(C and D) Knockdown of Beclin1 in MEFs by shRNA inhibits HBSS-induced autophagy with and without pUNO1-HMGB1 transfection. Cells as indicated were stimulated with Earle's balanced salt solution for 3 h, and LC3 expression was detected by Western blotting (C). | [
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F. PKCB forms an incoherent feed-forward loop with two parallel pathways, one suppressing fusion through VAMP8 phosphorylation and one promoting docking through putative phosphorylation substrates. | [
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(C) Knockdown efficiency of Rnf114 in the 2-cell stage embryos derived from Rnf114 siRNA #1 or siRNA #2 injected zygotes was verified by realtime-PCR and western blotting. NC indicates negative control injected with non-silencing siRNA. Three independent experiment replicates were performed for realtime-PCR, error bars represent s.e.m.; **:P<0.01, ***:P<0.01 in unpaired two-tailed t-test. | [
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Miniature inhibitory postsynaptic currents (mIPSCs) measured in CA1 pyramidal neurons in different mouse models at P14-P16. (A) example mIPSCs of WT (left) and Clcn3-/- neurons. | [
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(a,b) Venn diagrams representing the number of up- and down-regulated DEGs in response to HS (HS, LAT) with and without SA187 compared to NHS and NHS+187. The histograms show enriched GO terms for unique up- and down-regulated HS DEGs when compared to NHS. | [
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(D) Treatment of SARS-CoV-2 (106 PFU) infected kidney organoids with hrsACE2 (10µg/ml) and remdesivir (4µM). | [
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RT-PCR analysis of RNAs extracted from ARS immunoprecipitation (RIP) samples (erro bars: mean ± SD; biological replicates n = 3). Eight complex-forming ARSs (DARS, EPRS, IARS, KARS, LARS, MARS, QARS and RARS) and GARS tagged with FLAG were overexpressed in HEK cells. ARS-RNA complexes were imminoprecipitated with anti-FLAG Magnetic Beads. The cell line constructed with PQCXIP vector (PQ) was used as a negative control. GAPDH was used as an internal control for qRT-PCR. | [
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