ac p53 lys 382  (Cell Signaling Technology Inc)

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

Journal: Frontiers in Pharmacology

Article Title: Inhibition of SIRT3 by a specific inhibitor induces cellular senescence and growth arrest of ovarian granulosa cell tumor via p53 and NF-κB axis

doi: 10.3389/fphar.2025.1608156

Figure Lengend Snippet: 77-39 exerts its function through the p53 and NF-κB signaling pathways. (A) GSEA analysis showing senescence and NF-κB-related gene sets were enriched in the 77-39 treated group compared to the untreated group. (B) WB analyses showing that cell cycle-related proteins, p53, and NF-κB signaling were activated in a dose-dependent manner after treatment with 77–39.

Article Snippet: Anti-ac-p53 (HW137) antibody was purchased from Signalway (China).

Techniques: Protein-Protein interactions

Journal: Frontiers in Pharmacology

Article Title: Inhibition of SIRT3 by a specific inhibitor induces cellular senescence and growth arrest of ovarian granulosa cell tumor via p53 and NF-κB axis

doi: 10.3389/fphar.2025.1608156

Figure Lengend Snippet: Silencing SIRT3 in GCT cells has the same effect as treating with 77–39. (A) WB analyses showing the knockdown efficacy of SIRT3 siRNA in KGN and COV434 cells. (B) EdU assay showing the effects of SIRT3 silence on cell proliferation in KGN and COV434 cells. (C) SA-β-gal staining was used to evaluate the effects of the silence of SIRT3 on the cellular senescence of KGN and COV434. (D) WB analyses showing that cell cycle-related proteins, p53, and NF-κB signaling were activated after silencing SIRT3 in KGN and COV434 cells.

Article Snippet: Anti-ac-p53 (HW137) antibody was purchased from Signalway (China).

Techniques: Knockdown, EdU Assay, Staining

Journal: Frontiers in Pharmacology

Article Title: Inhibition of SIRT3 by a specific inhibitor induces cellular senescence and growth arrest of ovarian granulosa cell tumor via p53 and NF-κB axis

doi: 10.3389/fphar.2025.1608156

Figure Lengend Snippet: Overexpressing SIRT3 in GCT cells reverses the inhibitory effect of 77–39 in GCT cells. (A) WB analyses showing the successful overexpression of SIRT3 in KGN and COV434 cells. (B) SA-β-gal staining was used to evaluate the effects of overexpression of SIRT3 on the cellular senescence of KGN and COV434. (C) EdU assay showing the effects of SIRT3 overexpression on cell proliferation in KGN and COV434 cells. (D) WB analyses showing that cell cycle-related proteins, p53, and NF-κB signaling were inactivated after overexpressing SIRT3 in KGN and COV434 cells.

Article Snippet: Anti-ac-p53 (HW137) antibody was purchased from Signalway (China).

Techniques: Over Expression, Staining, EdU Assay

Journal: Frontiers in Pharmacology

Article Title: Inhibition of SIRT3 by a specific inhibitor induces cellular senescence and growth arrest of ovarian granulosa cell tumor via p53 and NF-κB axis

doi: 10.3389/fphar.2025.1608156

Figure Lengend Snippet: Inhibition of p53 counteracts the effect of 77–39 in GCT cells. (A) co-IP and WB analyses showing the interaction between SIRT3 and p53 in KGN and COV434 cells. (B) SA-β-gal staining was used to evaluate the effects of inhibition of p53 on the cellular senescence of KGN and COV434. (C) EdU assay showing the effects of p53 inhibition on cell proliferation in KGN and COV434 cells. (D) WB analyses showing that cell cycle-related proteins, p53, and NF-κB signaling were downregulated after p53 inhibition in KGN and COV434 cells.

Article Snippet: Anti-ac-p53 (HW137) antibody was purchased from Signalway (China).

Techniques: Inhibition, Co-Immunoprecipitation Assay, Staining, EdU Assay

Journal: Frontiers in Pharmacology

Article Title: Inhibition of SIRT3 by a specific inhibitor induces cellular senescence and growth arrest of ovarian granulosa cell tumor via p53 and NF-κB axis

doi: 10.3389/fphar.2025.1608156

Figure Lengend Snippet: Inhibition of NF-κB counteracts the effect of 77–39 in GCT cells. (A) SA-β-gal staining was used to evaluate the effects of inhibition of NF-κB on the cellular senescence of KGN and COV434. (B) EdU assay showing the effects of NF-κB inhibition on cell proliferation in KGN and COV434 cells. (C) WB analyses showing that cell cycle-related proteins, p53, and NF-κB signaling were downregulated after NF-κB inhibition in KGN and COV434 cells.

Article Snippet: Anti-ac-p53 (HW137) antibody was purchased from Signalway (China).

Techniques: Inhibition, Staining, EdU Assay

Journal: Frontiers in Pharmacology

Article Title: Inhibition of SIRT3 by a specific inhibitor induces cellular senescence and growth arrest of ovarian granulosa cell tumor via p53 and NF-κB axis

doi: 10.3389/fphar.2025.1608156

Figure Lengend Snippet: 77–39 suppresses GCT tumorigenicity in vivo . (A) Image of the subcutaneous KGN xenografts from mice that received indicated treatment. (B) Growth curves of KGN xenografts measured by a digital caliper. (C) Weight of KGN xenografts. (D) Body weight of mice that received indicated treatment. (E) KGN xenografts from the control and 77-39 (10 mg/kg) treated groups were subjected to HE staining, SA-β-gal staining, and IHC staining for Ki67, p21, p16, and p53. Histogram showing the statistical results and differences of the proportion of SA-β-gal positive cells and H-score. (F) Measurement of liver and kidney function markers in different treatment groups.

Article Snippet: Anti-ac-p53 (HW137) antibody was purchased from Signalway (China).

Techniques: In Vivo, Control, Staining, Immunohistochemistry

Journal: Aging Cell

Article Title: Targeting IGF1 ‐Induced Cellular Senescence to Rejuvenate Hair Follicle Aging

doi: 10.1111/acel.70053

Figure Lengend Snippet: Elevated IGF‐1 expression accelerates hair follicle aging through p53‐dependent cellular senescence. (a) Protein expressions of IGF‐1 in mouse skin are assessed by immunohistochemistry (IHC) and show a significant increase in aged mice. Representative staining and average optical density (AOD) quantification were demonstrated. Data are presented as means ± SEM, Two‐tailed Student's t ‐test. (b) IHC was performed on human perineal skin to measure IGF‐1 expression in two age groups: Younger women (< 30 years) and older women (> 60 years). Representative staining and AOD quantification were demonstrated. AOD analysis was performed on a minimum of 200 cells from the dorsal skin. Data are presented as means ± SEM, Two‐tailed Student's t ‐test. (c) Kaplan–Meier plot shows the lifespan follow‐up in male IGF‐1 Tg mice ( n = 36, red) or WT littermates ( n = 19, blue). p values were determined by the log‐rank (Mantel‐Cox) test. (d and e) Clinical frailty index scores and rotarod test of 10‐month WT and IGF‐1 Tg male mice ( n = 8/group). Data were means ± SEM. Statistical analysis was performed using a two‐tailed Student's t ‐test. (f) Representative images of dorsal skin coat color from 10‐month‐old IGF‐1 Tg mice, 10‐month‐old WT mice, and 24‐month‐old WT mice are shown. The age‐dependent appearance of gray hair in mice is graphed and quantified. (g) 10‐month‐old IGF‐1 Tg male mice and age‐matched WT male mice were shaved and monitored for hair coat recovery. Hair regrowth was quantified as the percentage of back skin covered by new hairs. Data are shown as means ± SD, n = 5/group. Two‐tailed Student's t ‐test. (h) Equal amounts of total tissue lysates from the dorsal skin of two 10‐month‐old IGF‐1 Tg male mice and their WT littermates were immunoprecipitated and analyzed by Western blotting to detect p53 and p21. The immunoprecipitated lysates were normalized for total p53, and the expression level of acetylated p53 at lysine 379 (Ac‐K379‐p53) was quantified. (i) Histological examination of dorsal skin was performed on 10‐month‐old IGF‐1 Tg mice, 10‐month‐old WT mice, and 24‐month‐old WT mice. Representative IHC images and quantitative analyses of IGF‐1, p53, p16, and PAI‐1 were presented. Data are shown as means ± SEM, n = 5/group. Two‐way ANOVA with Tukey's test. Scale bar = 50 μm.

Article Snippet: Antibodies for Ac‐K379‐p53 (#2570S, WB 1:1000), γ‐H2AX (#2577, WB 1:1000, IF 1:200), and PAI‐1 (#11907, WB 1:1000, IHC 1:200) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Immunohistochemistry, Staining, Two Tailed Test, Immunoprecipitation, Western Blot

Journal: Aging Cell

Article Title: Targeting IGF1 ‐Induced Cellular Senescence to Rejuvenate Hair Follicle Aging

doi: 10.1111/acel.70053

Figure Lengend Snippet: Forced SIRT1 overexpression attenuates hair graying and hair follicle cell senescence in IGF‐1 Tg mice. (a) Representative images of age‐associated coat color from 10‐month‐old male IGF‐1 Tg mice, SIRT1 Tg mice, IGF‐1/SIRT1 double transgenic (DTg) mice, and WT littermates, showing dorsal gray hair ( n = 10/group). The percentage of mice with gray hair in each group is also presented. (b) Equal amounts of total tissue lysates from the dorsal skin of these mice were immunoprecipitated and analyzed by Western blotting with a p53‐specific antibody. The immunoprecipitated samples were normalized to total p53 levels before immunoblotting for Ac‐K379‐p53. (c) Histological examination of dorsal skin was conducted to assess IGF‐1 signaling. Representative IHC images and corresponding quantitative analyses are shown for IGF‐1, SIRT1, p53, p16, and PAI‐1 ( n = 8/group). (d–g) Representative IF images and corresponding quantitative analyses of senescence‐associated DNA damage marker γ‐H2AX, SASP factor IL‐6, HFSC marker CD34, and melanocyte stem cell marker KIT ( n = 5/group). Data are presented as means ± SEM; two‐way ANOVA with Tukey's test. Scale bar = 50 μm.

Article Snippet: Antibodies for Ac‐K379‐p53 (#2570S, WB 1:1000), γ‐H2AX (#2577, WB 1:1000, IF 1:200), and PAI‐1 (#11907, WB 1:1000, IHC 1:200) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Over Expression, Transgenic Assay, Immunoprecipitation, Western Blot, Marker

Journal: Aging Cell

Article Title: Targeting IGF1 ‐Induced Cellular Senescence to Rejuvenate Hair Follicle Aging

doi: 10.1111/acel.70053

Figure Lengend Snippet: Dietary Restriction targeting IGF‐1 alleviates hair graying and HFSC senescence. (a) Four‐month‐old male IGF‐1 Tg mice and WT littermates were fed either regular chow or subjected to a 30% dietary restriction ( n = 11/group) for 6 months. Dorsal coat color and hair graying were assessed, followed by IHC and IF analyses of hair follicles. (b) Representative images of dorsal coat color and percentages of mice with gray hair in each group were analyzed. (c) Representative IHC images and corresponding quantitative analyses of IGF‐1, SIRT1, p53, p16, and PAI‐1 ( n = 7/group). (d–g) Representative IF images and corresponding quantitative analyses of the senescence‐associated DNA damage marker γ‐H2AX (d). SASP factor IL‐6 (e), hair follicle stem cell marker CD34 (f), and melanocyte stem cell marker KIT (g) ( n = 5/group). Data were means ± SEM, Two‐way ANOVA with Tukey's test. Scale bar = 50 μm. (h–j) 4‐month‐old male IGF‐1 Tg mice and WT littermates were fed either regular chow or subjected to a 30% dietary restriction ( n = 5/group) for 20 months. Dorsal coat color and hair graying were assessed, followed by IHC and IF analyses of hair follicles (h). Representative coat color images and AOD quantification are shown ( n = 5/group) (i). Representative IHC images and corresponding quantitative analyses of p53, p16, and PAI‐1 ( n = 5/group) (j).

Article Snippet: Antibodies for Ac‐K379‐p53 (#2570S, WB 1:1000), γ‐H2AX (#2577, WB 1:1000, IF 1:200), and PAI‐1 (#11907, WB 1:1000, IHC 1:200) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Marker