Journal: bioRxiv

Article Title: ATGL-catalyzed lipid catabolism promotes DNA repair

doi: 10.64898/2026.04.03.716381

Figure Lengend Snippet: A) Immunoblotting of pan-acetyl lysine from mouse livers. B) Immunoblot for γH2Ax and pan-acetyl lysine on chromatin fractions isolated from GFP- or ATGL-overexpressing AML12 cells. Cells were incubated in etoposide for 3 hours (“etop acute”) or allowed to recover for 3 hours after etoposide treatment (“etop recov”). C) Primary MEFs were pre-treated for 12 hours with various HAT inhibitors. Cells were then incubated in etoposide + inhibitors for 6 hours. Immunofluorescence was performed for γH2Ax, and cells were imaged using the Agilent Gen5 Cytation. CPTH2 = Gcn5 inhibitor, Nu9056 = Tip60 inhibitor, WM1119 = KAT6A inhibitor, A485 = p300 inhibitor. Quantification of γH2Ax positive cells (>2 foci per nucleus). Statistics: one-way ANOVA with Tukey’s post hoc test. Error bars indicate mean +/-SEM. *=p<0.05, **=p<0.01, ****=p<0.0001. D) Pathway analysis of AKI versus WT with differentially expressed genes (FDR 0.2) was performed with the TRRUST database. Top pathways shown with NES (normalized enrichment score). E) Representative images of p53 and ac-p53 (K379) immunofluorescence in AML12 cells overexpressing ATGL or Null control. Quantification of p53 and ac-p53 nuclear intensity (left) and nuclear intensity/cytoplasmic intensity (right). F) RT-qPCR of p53 target genes in WT and AKI Primary MEFs treated with etoposide for 3 hours, followed by 6 hours of recovery. G) AML12 cells were transduced with ATGL or a Null virus control. Cells were treated with A485 for 12 hours, followed by 60 minutes of etoposide treatment, followed by immunostaining for p53. *=p<0.05, **p<0.01. H) Primary MEFs were transfected with a pooled p53 siRNA and treated with etoposide for 3 hours, followed by a 1-hour recovery, and assayed for p53 and γH2Ax.

Article Snippet: For p53/ac-p53 imaging, primary antibodies (p53, Cell Signaling 2524, 1:300; Acetyl-p53, PA5-17287, 1:250) were diluted in PBS-T containing 5% goat serum and incubated with the sample for 2 hours at room temperature.

Techniques: Western Blot, Isolation, Incubation, Immunofluorescence, Control, Quantitative RT-PCR, Transduction, Virus, Immunostaining, Transfection

Journal: Cell Death Discovery

Article Title: ATGL sensitizes hepatocellular carcinoma cells to genotoxic drugs by modulating p53 acetylation/phosphorylation status

doi: 10.1038/s41420-026-03048-4

Figure Lengend Snippet: A , B HUH7 and C , D Hep3B cells were transfected with an empty vector (Vehicle) or ATGL-overexpressing construct (ATGL-OE) for 24 h and then treated with 50 µM etoposide or 2 µM doxorubicin for 6 h. Western blot analysis of γH2AX levels was performed. E Western blot analysis of Ac-p53 Lys-382, p-p53 Ser-15 and p53 levels was performed in HepG2 cells after treatment with 50 µM etoposide for 2, 4, 6 and 24 h. F Densitometric ratios of Ac-p53 and p-p53 after treatment with 50 µM etoposide for 2, 4, 6 and 24 h. G – I HepG2 cells were transfected with empty vector (Vehicle) or ATGL-overexpressing (ATGL-OE) construct and, after 24 h, treated with 50 µM etoposide for 6 h with or without 10 µM C646 for 24 h. Western blot analysis of Ac-p53 Lys-382, p-p53 Ser-15 and p53 levels was performed. H Densitometric analysis ratios of Ac-p53 and p-p53 after treatment with 50 µM etoposide for 6 h. J – L HepG2 cells were treated with 50 µM etoposide for 6 h with or without 1 µM GW7647 for 24 h. Western blot analysis of Ac-p53 Lys-382, p-p53 Ser-15, p53 and γH2AX levels was performed. K Densitometric analysis of ratio between Ac-p53 and p-p53 expression after treatment with 50 µM etoposide for 6 h. The images are representative of three independent experiments that yielded similar results. β-Actin and ATGL were used as loading and transfection controls, respectively. The data are presented as the means ± SDs from three independent experiments. Statistical significance was determined by Student t test and one-way ANOVA with Tukey’s post hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001 vs CTRL or as indicated by brackets.

Article Snippet: The following primary antibodies were used: β-Actin (Cell Signaling Technology, cat. number #4970S, diluted 1:1000), γH2AX Ser-139 (Cell Signaling Technology, cat. number #9718, diluted 1:1000), ATGL (Cell Signaling Technology, cat. number 2138S, diluted 1:1000), pATM Ser-1981 (Cell Signaling Technology, cat. number #5883, diluted 1:1000), ATM (Cell Signaling Technology, cat. number #2873, diluted 1:1000), p21 (Cell Signaling Technology, cat. number #2947, diluted 1:1000), PPARα (Santa Cruz Biotechnology, cat. number sc-398394, diluted 1:1000), Puma (Cell Signaling Technology, cat. number #4976, diluted 1:1000), Ac-p53 Lys-382 (Cell Signaling Technology, cat. number #2525S, diluted 1:1000), p-p53 Ser-15 (Cell Signaling Technology, cat. number #9284S, diluted 1:1000), and p53 (Sigma-Aldrich, cat. number #P5813, diluted 1:1000).

Techniques: Transfection, Plasmid Preparation, Construct, Western Blot, Expressing

Journal: Cell Death Discovery

Article Title: ATGL sensitizes hepatocellular carcinoma cells to genotoxic drugs by modulating p53 acetylation/phosphorylation status

doi: 10.1038/s41420-026-03048-4

Figure Lengend Snippet: A – D HepG2 cells were transfected with empty vector (Vehicle) or ATGL-overexpressing construct (ATGL-OE) and, after 24 h, treated with 50 µM etoposide for 6 h with or without recovery (Rec) with fresh medium for 2 h. Western blot analysis of Ac-p53 Lys-382, p-p53 Ser-15, p53 and γH2AX levels was performed. E , F HepG2 cells were transfected with empty vector (Vehicle) or ATGL-overexpressing construct (ATGL-OE) and, after 24 h, treated with 50 µM etoposide or 2 µM doxorubicin for 6 h. The proliferation was assayed via the Trypan blue direct counting procedure. G , H HepG2 cells were treated with 50 µM etoposide for 6 h with or without 25 µM ATGListatin (ATGLi) for 24 h. HepG2 cells were treated with 50 µM etoposide for 6 h. I – K Western blot analysis of p21 and Puma levels was performed. HepG2 cells were treated with 1 µM GW7647 for 24 h. L , M Proliferation was assayed by the Trypan blue direct counting procedure. N – P Western blot analysis of p21 and Puma levels was performed. The images are representative of three independent experiments that yielded similar results. β-Actin and ATGL were used as loading and transfection controls, respectively. The data are presented as the means ± SDs from three independent experiments. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001 vs CTRL or as indicated by brackets.

Article Snippet: The following primary antibodies were used: β-Actin (Cell Signaling Technology, cat. number #4970S, diluted 1:1000), γH2AX Ser-139 (Cell Signaling Technology, cat. number #9718, diluted 1:1000), ATGL (Cell Signaling Technology, cat. number 2138S, diluted 1:1000), pATM Ser-1981 (Cell Signaling Technology, cat. number #5883, diluted 1:1000), ATM (Cell Signaling Technology, cat. number #2873, diluted 1:1000), p21 (Cell Signaling Technology, cat. number #2947, diluted 1:1000), PPARα (Santa Cruz Biotechnology, cat. number sc-398394, diluted 1:1000), Puma (Cell Signaling Technology, cat. number #4976, diluted 1:1000), Ac-p53 Lys-382 (Cell Signaling Technology, cat. number #2525S, diluted 1:1000), p-p53 Ser-15 (Cell Signaling Technology, cat. number #9284S, diluted 1:1000), and p53 (Sigma-Aldrich, cat. number #P5813, diluted 1:1000).

Techniques: Transfection, Plasmid Preparation, Construct, Western Blot

Journal: Cell Death Discovery

Article Title: ATGL sensitizes hepatocellular carcinoma cells to genotoxic drugs by modulating p53 acetylation/phosphorylation status

doi: 10.1038/s41420-026-03048-4

Figure Lengend Snippet: A Boxplot showing significantly reduced PNPLA2 expression in primary HCC tissues (primary tumor; n = 371) compared with solid tumor-adjacent non-tumoral liver tissues (solid tissue normal; n = 50) samples based on TCGA data. B Boxplot of Z-score–normalized ATGL expression from TCGA-LIHC RNA-seq data based on TP53 mutation status (wild type n = 263 and mutant n = 111). C Visualization of the PPAR signaling pathway, reporting normalized enrichment score (NES) and adjusted p -value. D Bar plot showing the most significantly enriched transcription factors after a transcription factor enrichment analysis performed using TRRUST transcription factors 2019 database on differentially expressed genes (DEGs) in ATGL-high versus ATGL-low HCC samples. Adjusted p -value was reported. Scatter plot showing the correlation between E PNPLA2 and PPARα ( PPARA ); between F PNPLA2 and EP300 ; between G PPARA and EP300 ; between H PNPLA2 and Puma ( BBC3 ); between I PNPLA2 and p21 ( CDKN1A ) mRNA expression levels in HCC samples from the TCGA-LIHC cohort analyzed using GEPIA. Gene expression values are reported as log2-transformed TPM. Each dot represents an individual tumor sample.

Article Snippet: The following primary antibodies were used: β-Actin (Cell Signaling Technology, cat. number #4970S, diluted 1:1000), γH2AX Ser-139 (Cell Signaling Technology, cat. number #9718, diluted 1:1000), ATGL (Cell Signaling Technology, cat. number 2138S, diluted 1:1000), pATM Ser-1981 (Cell Signaling Technology, cat. number #5883, diluted 1:1000), ATM (Cell Signaling Technology, cat. number #2873, diluted 1:1000), p21 (Cell Signaling Technology, cat. number #2947, diluted 1:1000), PPARα (Santa Cruz Biotechnology, cat. number sc-398394, diluted 1:1000), Puma (Cell Signaling Technology, cat. number #4976, diluted 1:1000), Ac-p53 Lys-382 (Cell Signaling Technology, cat. number #2525S, diluted 1:1000), p-p53 Ser-15 (Cell Signaling Technology, cat. number #9284S, diluted 1:1000), and p53 (Sigma-Aldrich, cat. number #P5813, diluted 1:1000).

Techniques: Expressing, RNA Sequencing, Mutagenesis, Gene Expression, Transformation Assay