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Obesity Primary Antibodies Against Atf3, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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The <t>p38‐ATF3</t> axis mediates resveratrol‐induced GDF15 expression. (A) Volcano plot from RNA‐seq analysis of MEF cells treated with resveratrol (10 μM), highlighting Atf3 as a significantly upregulated transcript (A). ATF3 mRNA (B) and protein (C) expression in resveratrol‐treated MEF cells. Gdf15 mRNA levels in MEF cells transfected with control or Atf3 ‐targeting siRNA, followed by resveratrol treatment (D). KEGG pathway enrichment analysis of differentially expressed genes from RNA‐seq data (E). Gdf15 mRNA expression in MEF cells treated with a p38 inhibitor (SB203580, 10 μM) (F). Levels of ATF3, GDF15, p38, and p‐p38 proteins in HepG2 cells treated with resveratrol, a p38 inhibitor, or a combination of both (G). Data are mean ± SEM; * p < 0.05.
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ABclonal Biotechnology antibodies against atf3
Expression and diagnostic significance of <t>ATF3</t> in pan-cancer. A ATF3 expression in normal and cancer samples analyzed with the GTEx and TCGA datasets. B Receiver operating characteristic (ROC) analysis evaluated the diagnostic performance of ATF3 across cancer types. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001
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(A-B) Volcano plots of whole-transcriptome differentially expressed genes (DEGs) identified by RNAseq in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 ( n =3) compared to control ( n =3). Dashed vertical lines reflect log 2 FC of −1 and 1. Dashed horizontal line reflects false discovery rate (FDR) of 0.05. Downregulated genes with log 2 FC≤-1 and -log 10 adjusted p-value above FDR threshold are denoted in blue. Upregulated genes log 2 FC≥1 and -log 10 adjusted p-value above FDR threshold are denoted in red. Raw RNAseq hit counts for ZNF423 in NF90.8 cells transfected with control siRNA ( n =3) or siZNF423 ( n =3). P-values represent unpaired, two-tailed t-tests between groups. (C) Venn diagram showing overlap of significantly downregulated DEGs common among ST88-14 and NF90.8 cell lines. Downregulated DEGs were defined as log 2 FC≥-1 and adjusted p-value of ≤0.05. (D) Dot plot of top enriched Hallmark gene signatures of shared downregulated genes (FDR≤0.05). (E) Venn diagram showing overlap of significantly upregulated DEGs common among ST88-14 and NF90.8 cell lines. Upregulated DEGs were defined as log 2 FC≥1 and adjusted p-value of ≤0.05. (F) Dot plot of top enriched Hallmark gene signatures of shared upregulated genes (FDR≤0.05). (G-H) Volcano plots of differentially expressed transcription factors and kinases in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 compared to control. (I-J) Immunoblots showing cleavage of PARP, <t>ATF3,</t> ATF4 and ZNF423 protein expression following siRNA-mediated depletion of ZNF423 ( n =3) compared to controls ( n =3). GAPDH serves as loading control. Protein expression was normalized to total protein and quantified using ImageJ. Error bars reflect SEM. P-values represent unpaired, two-tailed t test with Welch’s correction. ****=p-value ≤0.0001, ***=p-value≤0.001, **=p-value≤0.01, *=p-value≤0.05, ns=not significant.
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(A-B) Volcano plots of whole-transcriptome differentially expressed genes (DEGs) identified by RNAseq in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 ( n =3) compared to control ( n =3). Dashed vertical lines reflect log 2 FC of −1 and 1. Dashed horizontal line reflects false discovery rate (FDR) of 0.05. Downregulated genes with log 2 FC≤-1 and -log 10 adjusted p-value above FDR threshold are denoted in blue. Upregulated genes log 2 FC≥1 and -log 10 adjusted p-value above FDR threshold are denoted in red. Raw RNAseq hit counts for ZNF423 in NF90.8 cells transfected with control siRNA ( n =3) or siZNF423 ( n =3). P-values represent unpaired, two-tailed t-tests between groups. (C) Venn diagram showing overlap of significantly downregulated DEGs common among ST88-14 and NF90.8 cell lines. Downregulated DEGs were defined as log 2 FC≥-1 and adjusted p-value of ≤0.05. (D) Dot plot of top enriched Hallmark gene signatures of shared downregulated genes (FDR≤0.05). (E) Venn diagram showing overlap of significantly upregulated DEGs common among ST88-14 and NF90.8 cell lines. Upregulated DEGs were defined as log 2 FC≥1 and adjusted p-value of ≤0.05. (F) Dot plot of top enriched Hallmark gene signatures of shared upregulated genes (FDR≤0.05). (G-H) Volcano plots of differentially expressed transcription factors and kinases in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 compared to control. (I-J) Immunoblots showing cleavage of PARP, <t>ATF3,</t> ATF4 and ZNF423 protein expression following siRNA-mediated depletion of ZNF423 ( n =3) compared to controls ( n =3). GAPDH serves as loading control. Protein expression was normalized to total protein and quantified using ImageJ. Error bars reflect SEM. P-values represent unpaired, two-tailed t test with Welch’s correction. ****=p-value ≤0.0001, ***=p-value≤0.001, **=p-value≤0.01, *=p-value≤0.05, ns=not significant.
Anti Atf3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A-B) Volcano plots of whole-transcriptome differentially expressed genes (DEGs) identified by RNAseq in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 ( n =3) compared to control ( n =3). Dashed vertical lines reflect log 2 FC of −1 and 1. Dashed horizontal line reflects false discovery rate (FDR) of 0.05. Downregulated genes with log 2 FC≤-1 and -log 10 adjusted p-value above FDR threshold are denoted in blue. Upregulated genes log 2 FC≥1 and -log 10 adjusted p-value above FDR threshold are denoted in red. Raw RNAseq hit counts for ZNF423 in NF90.8 cells transfected with control siRNA ( n =3) or siZNF423 ( n =3). P-values represent unpaired, two-tailed t-tests between groups. (C) Venn diagram showing overlap of significantly downregulated DEGs common among ST88-14 and NF90.8 cell lines. Downregulated DEGs were defined as log 2 FC≥-1 and adjusted p-value of ≤0.05. (D) Dot plot of top enriched Hallmark gene signatures of shared downregulated genes (FDR≤0.05). (E) Venn diagram showing overlap of significantly upregulated DEGs common among ST88-14 and NF90.8 cell lines. Upregulated DEGs were defined as log 2 FC≥1 and adjusted p-value of ≤0.05. (F) Dot plot of top enriched Hallmark gene signatures of shared upregulated genes (FDR≤0.05). (G-H) Volcano plots of differentially expressed transcription factors and kinases in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 compared to control. (I-J) Immunoblots showing cleavage of PARP, <t>ATF3,</t> ATF4 and ZNF423 protein expression following siRNA-mediated depletion of ZNF423 ( n =3) compared to controls ( n =3). GAPDH serves as loading control. Protein expression was normalized to total protein and quantified using ImageJ. Error bars reflect SEM. P-values represent unpaired, two-tailed t test with Welch’s correction. ****=p-value ≤0.0001, ***=p-value≤0.001, **=p-value≤0.01, *=p-value≤0.05, ns=not significant.
Anti Atf3, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


The p38‐ATF3 axis mediates resveratrol‐induced GDF15 expression. (A) Volcano plot from RNA‐seq analysis of MEF cells treated with resveratrol (10 μM), highlighting Atf3 as a significantly upregulated transcript (A). ATF3 mRNA (B) and protein (C) expression in resveratrol‐treated MEF cells. Gdf15 mRNA levels in MEF cells transfected with control or Atf3 ‐targeting siRNA, followed by resveratrol treatment (D). KEGG pathway enrichment analysis of differentially expressed genes from RNA‐seq data (E). Gdf15 mRNA expression in MEF cells treated with a p38 inhibitor (SB203580, 10 μM) (F). Levels of ATF3, GDF15, p38, and p‐p38 proteins in HepG2 cells treated with resveratrol, a p38 inhibitor, or a combination of both (G). Data are mean ± SEM; * p < 0.05.

Journal: Food Science & Nutrition

Article Title: Resveratrol Attenuates Obesity by Inducing GDF15 Expression via the p38‐ ATF3 Signaling Pathway

doi: 10.1002/fsn3.72052

Figure Lengend Snippet: The p38‐ATF3 axis mediates resveratrol‐induced GDF15 expression. (A) Volcano plot from RNA‐seq analysis of MEF cells treated with resveratrol (10 μM), highlighting Atf3 as a significantly upregulated transcript (A). ATF3 mRNA (B) and protein (C) expression in resveratrol‐treated MEF cells. Gdf15 mRNA levels in MEF cells transfected with control or Atf3 ‐targeting siRNA, followed by resveratrol treatment (D). KEGG pathway enrichment analysis of differentially expressed genes from RNA‐seq data (E). Gdf15 mRNA expression in MEF cells treated with a p38 inhibitor (SB203580, 10 μM) (F). Levels of ATF3, GDF15, p38, and p‐p38 proteins in HepG2 cells treated with resveratrol, a p38 inhibitor, or a combination of both (G). Data are mean ± SEM; * p < 0.05.

Article Snippet: Equal amounts of protein were separated by SDS‐PAGE, transferred to PVDF membranes, and probed with specific primary antibodies against GDF15 (Abclonal, A0185), ATF3 (Abclonal, A1852), P38MAPK (Abclonal, A4771), Phospho‐P38MAPK (Abclonal, AP1372), phospho‐ERK (CST, 4370), total ERK (CST, 4695), GAPDH (CST, 5174 T) and beta‐actin (ACTB) (Abclonal, AC026).

Techniques: Expressing, RNA Sequencing, Transfection, Control

Tissue induction of ATF3 and GDF15 by resveratrol in vivo. (A, B) Relative mRNA expression of Atf3 (A) and Gdf15 (B) in liver, kidney, and colon tissues from resveratrol‐treated mice. (C–E) Representative Western blots and quantification of GDF15 protein expression in kidney (C), liver (D), and colon (E) tissues. Data are mean ± SEM; * p < 0.05.

Journal: Food Science & Nutrition

Article Title: Resveratrol Attenuates Obesity by Inducing GDF15 Expression via the p38‐ ATF3 Signaling Pathway

doi: 10.1002/fsn3.72052

Figure Lengend Snippet: Tissue induction of ATF3 and GDF15 by resveratrol in vivo. (A, B) Relative mRNA expression of Atf3 (A) and Gdf15 (B) in liver, kidney, and colon tissues from resveratrol‐treated mice. (C–E) Representative Western blots and quantification of GDF15 protein expression in kidney (C), liver (D), and colon (E) tissues. Data are mean ± SEM; * p < 0.05.

Article Snippet: Equal amounts of protein were separated by SDS‐PAGE, transferred to PVDF membranes, and probed with specific primary antibodies against GDF15 (Abclonal, A0185), ATF3 (Abclonal, A1852), P38MAPK (Abclonal, A4771), Phospho‐P38MAPK (Abclonal, AP1372), phospho‐ERK (CST, 4370), total ERK (CST, 4695), GAPDH (CST, 5174 T) and beta‐actin (ACTB) (Abclonal, AC026).

Techniques: In Vivo, Expressing, Western Blot

Proposed model for resveratrol‐induced body weight reduction via the GDF15 pathway. Schematic illustration summarizing the proposed mechanism: Resveratrol activates the p38 MAPK signaling pathway, leading to upregulation of ATF3. This transcription factor then promotes GDF15 expression and secretion in peripheral tissues. The elevated circulating GDF15 acts on the brain, potentially through activation of the ERK pathway in the hypothalamus, resulting in reduced food intake and subsequent protection against diet‐induced obesity.

Journal: Food Science & Nutrition

Article Title: Resveratrol Attenuates Obesity by Inducing GDF15 Expression via the p38‐ ATF3 Signaling Pathway

doi: 10.1002/fsn3.72052

Figure Lengend Snippet: Proposed model for resveratrol‐induced body weight reduction via the GDF15 pathway. Schematic illustration summarizing the proposed mechanism: Resveratrol activates the p38 MAPK signaling pathway, leading to upregulation of ATF3. This transcription factor then promotes GDF15 expression and secretion in peripheral tissues. The elevated circulating GDF15 acts on the brain, potentially through activation of the ERK pathway in the hypothalamus, resulting in reduced food intake and subsequent protection against diet‐induced obesity.

Article Snippet: Equal amounts of protein were separated by SDS‐PAGE, transferred to PVDF membranes, and probed with specific primary antibodies against GDF15 (Abclonal, A0185), ATF3 (Abclonal, A1852), P38MAPK (Abclonal, A4771), Phospho‐P38MAPK (Abclonal, AP1372), phospho‐ERK (CST, 4370), total ERK (CST, 4695), GAPDH (CST, 5174 T) and beta‐actin (ACTB) (Abclonal, AC026).

Techniques: Expressing, Activation Assay

Expression and diagnostic significance of ATF3 in pan-cancer. A ATF3 expression in normal and cancer samples analyzed with the GTEx and TCGA datasets. B Receiver operating characteristic (ROC) analysis evaluated the diagnostic performance of ATF3 across cancer types. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Expression and diagnostic significance of ATF3 in pan-cancer. A ATF3 expression in normal and cancer samples analyzed with the GTEx and TCGA datasets. B Receiver operating characteristic (ROC) analysis evaluated the diagnostic performance of ATF3 across cancer types. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques: Expressing, Diagnostic Assay

Prognostic evaluation of ATF3 in pan-cancer. A – D Univariate Cox regression analyses of ATF3 in different tumors for disease-free interval (DFI) ( A ), disease-specific survival (DSS) ( B ), overall survival (OS) ( C ) and progression-free interval (PFI) ( D ). E – H Patients were stratified by the median ATF3 expression; Kaplan-Meier curves compared the impact of high vs. low ATF3 expression on DFI ( E ), DSS ( F ), OS ( G ) and PFI ( H ) in each cancer type. HR, hazard ratio; CI, confidence interval

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Prognostic evaluation of ATF3 in pan-cancer. A – D Univariate Cox regression analyses of ATF3 in different tumors for disease-free interval (DFI) ( A ), disease-specific survival (DSS) ( B ), overall survival (OS) ( C ) and progression-free interval (PFI) ( D ). E – H Patients were stratified by the median ATF3 expression; Kaplan-Meier curves compared the impact of high vs. low ATF3 expression on DFI ( E ), DSS ( F ), OS ( G ) and PFI ( H ) in each cancer type. HR, hazard ratio; CI, confidence interval

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques: Expressing

Mutation landscape and mutation-load analysis of ATF3 in pan-cancer. A Frequencies of distinct ATF3 mutation types across cancers. B Analysis showing that missense mutations (green) were the predominant ATF3 alteration. C Radar plot depicting the correlation between ATF3 expression and tumor mutation burden (TMB). D Radar plot depicting the correlation between ATF3 expression and microsatellite instability (MSI). *, P < 0.05; **, P < 0.01

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Mutation landscape and mutation-load analysis of ATF3 in pan-cancer. A Frequencies of distinct ATF3 mutation types across cancers. B Analysis showing that missense mutations (green) were the predominant ATF3 alteration. C Radar plot depicting the correlation between ATF3 expression and tumor mutation burden (TMB). D Radar plot depicting the correlation between ATF3 expression and microsatellite instability (MSI). *, P < 0.05; **, P < 0.01

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques: Mutagenesis, Expressing

Relationship between ATF3 expression and immunity/differentially methylated probes-based stemness index (DMPsi). A ESTIMATE analysis of the association between ATF3 expression and ImmuneScore, StromalScore, and EstimateScore. B – F Correlations of ATF3 expression with immune-checkpoint genes ( B ), chemokines (C), immune inhibitors (D), immune stimulators (E) and chemokines receptors (F). G Correlations between ATF3 and mismatch-repair and DNA-methyltransferase genes. H Correlation between ATF3 and DMPsi. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Relationship between ATF3 expression and immunity/differentially methylated probes-based stemness index (DMPsi). A ESTIMATE analysis of the association between ATF3 expression and ImmuneScore, StromalScore, and EstimateScore. B – F Correlations of ATF3 expression with immune-checkpoint genes ( B ), chemokines (C), immune inhibitors (D), immune stimulators (E) and chemokines receptors (F). G Correlations between ATF3 and mismatch-repair and DNA-methyltransferase genes. H Correlation between ATF3 and DMPsi. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques: Expressing, Methylation

ATF3 was down-regulated in kidney clear cell renal cell carcinoma (KIRC) and associated with favorable prognosis. A – C ATF3 expression in KIRC cell lines determined by real-time quantitative polymerase chain reaction (RT-qPCR) ( A ), Western blot ( B ) and immunofluorescence ( C ). D Kaplan-Meier survival curves comparing high vs. low ATF3 expression for DFI, DSS, OS and PFI in KIRC. E Association between ATF3 and clinical-feature subgroups in KIRC. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: ATF3 was down-regulated in kidney clear cell renal cell carcinoma (KIRC) and associated with favorable prognosis. A – C ATF3 expression in KIRC cell lines determined by real-time quantitative polymerase chain reaction (RT-qPCR) ( A ), Western blot ( B ) and immunofluorescence ( C ). D Kaplan-Meier survival curves comparing high vs. low ATF3 expression for DFI, DSS, OS and PFI in KIRC. E Association between ATF3 and clinical-feature subgroups in KIRC. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Immunofluorescence

Enrichment and immune-infiltration analyses of ATF3. A Volcano plot of differentially expressed genes (DEGs) between high and low ATF3 groups. B Heat-map of the DEGs. C Gene Ontology (GO) enrichment of the DEGs. BP, Biological Process; CC, Cellular Component; MF, Molecular Function. D Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of the DEGs. E Gene set enrichment analysis (GSEA) of ATF3. F Stacked bar chart of immune-cell scores in high and low ATF3 groups. G Box plot showing differences in immune-cell infiltration between the two groups. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Enrichment and immune-infiltration analyses of ATF3. A Volcano plot of differentially expressed genes (DEGs) between high and low ATF3 groups. B Heat-map of the DEGs. C Gene Ontology (GO) enrichment of the DEGs. BP, Biological Process; CC, Cellular Component; MF, Molecular Function. D Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of the DEGs. E Gene set enrichment analysis (GSEA) of ATF3. F Stacked bar chart of immune-cell scores in high and low ATF3 groups. G Box plot showing differences in immune-cell infiltration between the two groups. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques:

Prediction of ATF3-targeted drugs. A Drug-sensitivity analysis between high and low ATF3 groups (top 10 agents). B ATF3-drug network. C Molecular-docking affinity between progesterone and ATF3

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Prediction of ATF3-targeted drugs. A Drug-sensitivity analysis between high and low ATF3 groups (top 10 agents). B ATF3-drug network. C Molecular-docking affinity between progesterone and ATF3

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques:

Single-cell RNA sequencing (scRNA-seq) analysis of ATF3 in KIRC. A Uniform Manifold Approximation and Projection (UMAP) showing cell annotations. B Proportions of annotated cells in KIRC vs. control. C ATF3 expression across cell types in KIRC vs. control. D Pseudotime trajectory of endothelial-afferent/efferent arterioles/descending vasa recta (AEAs/DVR) divided into three stages. E Distribution of KIRC and control samples along the AEAs/DVR differentiation trajectory. F ATF3 expression during AEAs/DVR differentiation. G Cell-cell communication analysis. *, P < 0.05; **, P < 0.01; ****, P < 0.0001

Journal: Discover Oncology

Article Title: Comprehensive analysis of ATF3 as a diagnostic and prognostic biomarker from pan-cancer to clear cell renal cell carcinoma

doi: 10.1007/s12672-026-05113-x

Figure Lengend Snippet: Single-cell RNA sequencing (scRNA-seq) analysis of ATF3 in KIRC. A Uniform Manifold Approximation and Projection (UMAP) showing cell annotations. B Proportions of annotated cells in KIRC vs. control. C ATF3 expression across cell types in KIRC vs. control. D Pseudotime trajectory of endothelial-afferent/efferent arterioles/descending vasa recta (AEAs/DVR) divided into three stages. E Distribution of KIRC and control samples along the AEAs/DVR differentiation trajectory. F ATF3 expression during AEAs/DVR differentiation. G Cell-cell communication analysis. *, P < 0.05; **, P < 0.01; ****, P < 0.0001

Article Snippet: After 30 min of blocking with 5% non-fat milk, the membranes were incubated for the whole night at 4 °C with primary antibodies against ATF3 (1:500, ABclonal) and GAPDH (1:10,000, ABclonal).

Techniques: Single Cell, RNA Sequencing, Control, Expressing

(A-B) Volcano plots of whole-transcriptome differentially expressed genes (DEGs) identified by RNAseq in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 ( n =3) compared to control ( n =3). Dashed vertical lines reflect log 2 FC of −1 and 1. Dashed horizontal line reflects false discovery rate (FDR) of 0.05. Downregulated genes with log 2 FC≤-1 and -log 10 adjusted p-value above FDR threshold are denoted in blue. Upregulated genes log 2 FC≥1 and -log 10 adjusted p-value above FDR threshold are denoted in red. Raw RNAseq hit counts for ZNF423 in NF90.8 cells transfected with control siRNA ( n =3) or siZNF423 ( n =3). P-values represent unpaired, two-tailed t-tests between groups. (C) Venn diagram showing overlap of significantly downregulated DEGs common among ST88-14 and NF90.8 cell lines. Downregulated DEGs were defined as log 2 FC≥-1 and adjusted p-value of ≤0.05. (D) Dot plot of top enriched Hallmark gene signatures of shared downregulated genes (FDR≤0.05). (E) Venn diagram showing overlap of significantly upregulated DEGs common among ST88-14 and NF90.8 cell lines. Upregulated DEGs were defined as log 2 FC≥1 and adjusted p-value of ≤0.05. (F) Dot plot of top enriched Hallmark gene signatures of shared upregulated genes (FDR≤0.05). (G-H) Volcano plots of differentially expressed transcription factors and kinases in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 compared to control. (I-J) Immunoblots showing cleavage of PARP, ATF3, ATF4 and ZNF423 protein expression following siRNA-mediated depletion of ZNF423 ( n =3) compared to controls ( n =3). GAPDH serves as loading control. Protein expression was normalized to total protein and quantified using ImageJ. Error bars reflect SEM. P-values represent unpaired, two-tailed t test with Welch’s correction. ****=p-value ≤0.0001, ***=p-value≤0.001, **=p-value≤0.01, *=p-value≤0.05, ns=not significant.

Journal: bioRxiv

Article Title: ZNF423 depletion induces the integrated stress response and represents a potential vulnerability in NF1-associated MPNST

doi: 10.64898/2026.03.03.709360

Figure Lengend Snippet: (A-B) Volcano plots of whole-transcriptome differentially expressed genes (DEGs) identified by RNAseq in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 ( n =3) compared to control ( n =3). Dashed vertical lines reflect log 2 FC of −1 and 1. Dashed horizontal line reflects false discovery rate (FDR) of 0.05. Downregulated genes with log 2 FC≤-1 and -log 10 adjusted p-value above FDR threshold are denoted in blue. Upregulated genes log 2 FC≥1 and -log 10 adjusted p-value above FDR threshold are denoted in red. Raw RNAseq hit counts for ZNF423 in NF90.8 cells transfected with control siRNA ( n =3) or siZNF423 ( n =3). P-values represent unpaired, two-tailed t-tests between groups. (C) Venn diagram showing overlap of significantly downregulated DEGs common among ST88-14 and NF90.8 cell lines. Downregulated DEGs were defined as log 2 FC≥-1 and adjusted p-value of ≤0.05. (D) Dot plot of top enriched Hallmark gene signatures of shared downregulated genes (FDR≤0.05). (E) Venn diagram showing overlap of significantly upregulated DEGs common among ST88-14 and NF90.8 cell lines. Upregulated DEGs were defined as log 2 FC≥1 and adjusted p-value of ≤0.05. (F) Dot plot of top enriched Hallmark gene signatures of shared upregulated genes (FDR≤0.05). (G-H) Volcano plots of differentially expressed transcription factors and kinases in ST88-14 and NF90.8 cells following siRNA-mediated depletion of ZNF423 compared to control. (I-J) Immunoblots showing cleavage of PARP, ATF3, ATF4 and ZNF423 protein expression following siRNA-mediated depletion of ZNF423 ( n =3) compared to controls ( n =3). GAPDH serves as loading control. Protein expression was normalized to total protein and quantified using ImageJ. Error bars reflect SEM. P-values represent unpaired, two-tailed t test with Welch’s correction. ****=p-value ≤0.0001, ***=p-value≤0.001, **=p-value≤0.01, *=p-value≤0.05, ns=not significant.

Article Snippet: Immunoblots were performed using primary antibodies against ZNF423 (ABN410, Sigma-Aldrich), GAPDH (sc-365062, Santa Cruz Biotechnology), SUZ12 (3737S, Cell Signaling Technology), ATF3 (18665S, Cell Signaling Technology), ATF4 (11815S, Cell Signaling Technology), PARP (9542S, Cell Signaling Technology), and γH2A.x (9718S, Cell Signaling Technology).

Techniques: RNA sequencing, Control, Transfection, Two Tailed Test, Western Blot, Expressing