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