anti acetyl p53  (Cell Signaling Technology Inc)

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk

doi: 10.3390/ijms27041841

Figure Lengend Snippet: Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and TRF2) proteins ( vi ).

Article Snippet: 4 , ACp53 , Cell Signaling Technology , 2570 , 1:1000 , .

Techniques: Saline, Immunohistochemistry, Isolation, Expressing, Western Blot, Staining, Control, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk

doi: 10.3390/ijms27041841

Figure Lengend Snippet: IL-17A induces p53, PAI-1, Cav1 and apoptosis, and CSP or CSP7 inhibits IL-17A-induced apoptosis in AECs. ( A ) AECs isolated from uninjured WT and p53 −/− mice were treated with IL-17A (0–100 ng/mL) for 24 h, and the lysates were immunoblotted for p53, PAI-1, Cl./Cas-3 and β-actin. ( B ) WT mice received saline or IL-17A intranasally (0–3 μg in 50 μL saline), as previously described . AECs were isolated 24 h later, and lysates were immunoblotted for p53 and Cl./Cas-3 and β-actin. ( C ) WT mice were exposed to saline or IL-17A (1 μg), as described in ( B ). Lysates of AECs isolated 24 h after treatment with IL-17A were immunoblotted for ACp53, p53, Sirt1, Cav1 and apoptosis. ( D ) AECs isolated from WT mice were treated with PBS or IL-17A (100 ng/mL). Two hours later, IL-17A-stimulated cells were treated with or without CSP or CP (10 µM) and analyzed for p53, PAI-1 and apoptosis by Western blotting (WB). ( E ) AECs isolated from WT mice exposed to IL-17A (100 ng/mL) were treated with CSP, its overlapping deletion fragments, or control peptide (CP) for 24 h in vitro. The lysates were analyzed for ACp53, p53, PAI-1, Sirt1 and Cl. Cas-3 by WB. ( F ) miR-34a fl/fl mice and miR-34acKO mice lacking miR-34a in AECs were exposed to IL-17A (1 μg) with or without CSP7 or CP. Control mice were exposed to saline. Lysates from AECs extracted from these mice were analyzed for p53, PAI-1, Sirt1, Cav1 and apoptosis by WB. ( G ) Lung homogenates or lysates of AECs extracted from WT mice exposed to IL-17A with or without CSP7 or CP were subjected to WB for PP2Ac, pATM and ATM kinase. ( H ) Lung homogenates of IL-17A-exposed mice treated with or without CSP7 or CP were immunoprecipitated (IP) using anti-Cav1 antibody and immunoblotted for p53, mdm2, and PP2Ac. ( I ) Lung homogenates of WT mice exposed to saline or IL-17A with or without CSP7 were analyzed for MPO activity.

Article Snippet: 4 , ACp53 , Cell Signaling Technology , 2570 , 1:1000 , .

Techniques: Isolation, Saline, Western Blot, Control, In Vitro, Immunoprecipitation, Activity Assay

Journal: Communications Biology

Article Title: The p53 R181C mutation accumulates through impaired deacetylation by Sirt1 and facilitates tumor development

doi: 10.1038/s42003-025-09465-y

Figure Lengend Snippet: A Common types and proportions of cancer in the population that are associated with the p53R181C mutation. B Expression levels of p53 protein in heart, liver, spleen, and kidney tissues of wild type and p53R178C mutant mice. C – F HEK293 cells were transfected with Flag-p53WT and Flag-p53R181C plasmids. Exogenous p53WT and p53R181C were analyzed for phosphorylation at the Ser15 site ( C ) and Ser20 site ( D ), and for levels of O-GlcNAc glycosylation ( E ), and acetylation ( F ). AC/Flag protein levels were determined by western blot analysis. Statistical significance of protein expression levels was assessed by Student’s t test (*P < 0.05; **P < 0.01; ***P < 0.001). G The acetylation level of the K382 site of Flag-p53WT and Flag-p53R181C was determined using a p53K382-specific antibody. Total protein lysates from HEK293 cells expressing Flag-p53WT or Flag-p53R181C were subjected to immunoprecipitation and immunoblotting to assess acetylation of K382. H Total protein lysates from NALM6 cells and REH cells were subjected immunoprecipitation and immunoblotting to assess the K382 acetylation of endogenous p53. NALM6 cells carry wild-type p53, whereas REH cells harbor the p53-R181C mutation. I Total protein lysates from wild-type and p53R178C mutant MEF cells were subjected immunoprecipitation and immunoblotting to assess the K382 acetylation of endogenous p53. J Acetylation-inactive mutations were introduced at the K382 site of Flag-p53WT and Flag-p53R181C to evaluate their acetylation levels. K The K382 acetylation levels of p53 in wild-type and p53R178C mutant MEF cells were examined during passage culture. L The K382 acetylation levels of p53 in wild-type and p53R178C mutant MEF cells were analyzed under DNA damage stress conditions with or without exposure to IR. γ-H2AX serves as a marker for DNA damage. M Total protein lysates from HEK293 p53-KO cells expressing Flag-Vector, Flag-WT, Flag-R181C, Flag-R175H, Flag-R248W, and Flag-R273H were subjected to western blot analysis of acetylation of K382.

Article Snippet: The antibodies used in this study include: anti-p53 (#sc-126, Santa Cruz, 1:1000), anti-p21 (#sc-6246, Santa Cruz, 1:1000), anti-Nedd8 (#sc-373741, Santa Cruz, 1:1000), anti-GFP (#A02375-50, Genscript, 1:2000), anti-Phospho-p53 (Ser20) (#9287S, Cell Signaling Technology, 1:1000), anti-Phospho-p53 (Ser15) (#9284S, Cell Signaling Technology, 1:1000), anti-Acetyl-p53 (Lys379) (#2570S, Cell Signaling Technology, 1:1000), anti-MDM2(#51541S, Cell Signaling Technology, 1:1000), anti-ATM(#2873S, Cell Signaling Technology, 1:1000), anti-Phospho-ATM (Ser1981) (#13050S, Cell Signaling Technology, 1:1000), anti-CHK2(#3440S, Cell Signaling Technology, 1:1000), anti-Phospho-CHK2 (Thr68) (#2661S, Cell Signaling Technology, 1:1000), anti-BCL2(#15071S, Cell Signaling Technology, 1:1000), anti-Phospho-Histone H2A.X (Ser139) (#9718S, Cell Signaling Technology, 1:1000), anti-acetylated-lysine (#9441S, Cell Signaling Technology, 1:1000), anti-HA (#3724, Cell Signaling Technology, 1:1000), anti-β-actin (AC004, ABclonal, 1:2000), anti-α-Tubulin (AC012, ABclonal, 1:2000), anti-Flag (#SG4110-16, Shanghai Genomics Technology, 1:2000), anti-myc (SG4110-18, Shanghai Genomics Technology, 1:2000), anti-GAPDH (A19056, ABclonal, 1:2000), anti-Bax (A19684, ABclonal, 1:1000), anti-PUMA (A3752, ABclonal, 1:1000), anti-p53(10442-1-AP, Proteintech, 1:1000), anti-Sirt1(#07-131, Millipore,1:1000).

Techniques: Mutagenesis, Expressing, Transfection, Phospho-proteomics, Glycoproteomics, Western Blot, Immunoprecipitation, Marker, Plasmid Preparation

Journal: Communications Biology

Article Title: The p53 R181C mutation accumulates through impaired deacetylation by Sirt1 and facilitates tumor development

doi: 10.1038/s42003-025-09465-y

Figure Lengend Snippet: A Analysis of Flag-p53WT and Flag-p53R181C binding to Sirt1. Lysates prepared from HEK293 p53KO cells transfected with Flag-p53WT or Flag-p53R181C were immunoprecipitated using anti-Sirt1 antibodies and the levels of co-precipitating Flag-p53WT and Flag-p53R181C were determined by immunoblotting with anti-Flag antibodies. B Lysates prepared as in ( A ) were immunoprecipitated using anti-Flag antibodies and the level of co-precipitating Sirt1 was detected with anti-Sirt1 antibodies. C Analysis of interaction of endogenous p53WT and p53R181C with Sirt1. Lysates from NALM6 and REH cells were subjected to immunoprecipitation with control IgG or anti-p53 antibody, followed by immunoblotting with anti-Sirt1 antibody. D Detection of the acetylation levels of Flag-p53WT and Flag-p53R181C at K382 under similar expression efficiency with or without Sirt1 knockout in HEK293 cells. The acetylation levels of Flag-p53WT and Flag-p53R181C at K382 in p53KO HEK293 cells were detected in the presence or absence of SIRT1 inhibitor EX527 ( E ), or under the condition of Sirt1 activation with the enzyme activator resveratrol ( F ). G H1299 cells were transfected with Flag-p53WT and Flag-p53R181C plasmids, respectively. After 48 h, the cells were treated with 200 µM H 2 O 2 for 4 h, and the acetylation level at p53 K382 was subsequently detected. The mRNA from third-generation wild type and p53R178C MEF cells was extracted followed by real-time fluorescence quantitative PCR analysis to detect the mRNA levels of endogenous p53WT and p53R178C under the conditions of Sirt1 deacetylase activity activation by resveratrol ( H ) or inhibition by EX527 ( I ).

Article Snippet: The antibodies used in this study include: anti-p53 (#sc-126, Santa Cruz, 1:1000), anti-p21 (#sc-6246, Santa Cruz, 1:1000), anti-Nedd8 (#sc-373741, Santa Cruz, 1:1000), anti-GFP (#A02375-50, Genscript, 1:2000), anti-Phospho-p53 (Ser20) (#9287S, Cell Signaling Technology, 1:1000), anti-Phospho-p53 (Ser15) (#9284S, Cell Signaling Technology, 1:1000), anti-Acetyl-p53 (Lys379) (#2570S, Cell Signaling Technology, 1:1000), anti-MDM2(#51541S, Cell Signaling Technology, 1:1000), anti-ATM(#2873S, Cell Signaling Technology, 1:1000), anti-Phospho-ATM (Ser1981) (#13050S, Cell Signaling Technology, 1:1000), anti-CHK2(#3440S, Cell Signaling Technology, 1:1000), anti-Phospho-CHK2 (Thr68) (#2661S, Cell Signaling Technology, 1:1000), anti-BCL2(#15071S, Cell Signaling Technology, 1:1000), anti-Phospho-Histone H2A.X (Ser139) (#9718S, Cell Signaling Technology, 1:1000), anti-acetylated-lysine (#9441S, Cell Signaling Technology, 1:1000), anti-HA (#3724, Cell Signaling Technology, 1:1000), anti-β-actin (AC004, ABclonal, 1:2000), anti-α-Tubulin (AC012, ABclonal, 1:2000), anti-Flag (#SG4110-16, Shanghai Genomics Technology, 1:2000), anti-myc (SG4110-18, Shanghai Genomics Technology, 1:2000), anti-GAPDH (A19056, ABclonal, 1:2000), anti-Bax (A19684, ABclonal, 1:1000), anti-PUMA (A3752, ABclonal, 1:1000), anti-p53(10442-1-AP, Proteintech, 1:1000), anti-Sirt1(#07-131, Millipore,1:1000).

Techniques: Binding Assay, Transfection, Immunoprecipitation, Western Blot, Control, Expressing, Knock-Out, Activation Assay, Fluorescence, Real-time Polymerase Chain Reaction, Histone Deacetylase Assay, Activity Assay, Inhibition

Journal: Communications Biology

Article Title: The p53 R181C mutation accumulates through impaired deacetylation by Sirt1 and facilitates tumor development

doi: 10.1038/s42003-025-09465-y

Figure Lengend Snippet: A Co-IP analysis using anti-Flag antibodies to detect endogenous MDM2 bound to Flag-p53WT and Flag-p53R181C in HEK293 p53KO cells. B Co-IP analysis using anti-MDM2 antibodies to detect Flag-p53WT and Flag-p53R181C bound to MDM2 in HEK293 p53KO cells. C Analysis of interaction of endogenous p53WT and p53R181C with MDM2. Lysates from NALM6 and REH cells were subjected to immunoprecipitation with control IgG or anti-p53 antibody, followed by immunoblotting with anti-MDM2 antibody. D Flag-p53WT or Flag-p53R181C was co-transfected with HA-Ub and myc-MDM2, followed by immunoprecipitation using anti-HA antibodies and detection of Flag by immunoblotting. E Analysis of endogenous ubiquitination levels of p53WT and p53R181C. Lysates from NALM6 and REH cells were immunoprecipitated with control IgG or anti-Ub antibody, followed by immunoblotting with anti-p53 antibody. F In vitro ubiquitination assay to determine the direct effect of MDM2 on the ubiquitination of p53WT and p53R181C. His-MDM2, GST-p53WT, and GST-p53R181C were purified in vitro. These purified proteins were incubated with Mg-ATP, Ubiquitin, E1, and E2 at 37 °C for 1 h. After termination, the reaction was subjected to immunoblotting with anti-ubiquitin antibody. G , H Co-IP analysis was performed to determine the effect of the different acetylation states (K382Q, K382R) on the interaction between MDM2 and Flag-p53R181C. I Detection of the ubiquitination levels of Flag-p53WT and Flag-p53R181C in HEK293 cells, with or without Sirt1 knockout. Lysates were subjected to immunoprecipitation with anti-HA antibody, followed by immunoblotting with anti-Flag antibody. Measurement of the ubiquitination levels of Flag-p53WT and Flag-p53R181C in p53KO HEK293 cells with activation of Sirt1 deacetylase activity by resveratrol ( J ) and inhibition of Sirt1 deacetylase activity by EX527( K ). Lysates were subjected to immunoprecipitation with anti-HA antibody, followed by immunoblotting with anti-Flag antibody.

Article Snippet: The antibodies used in this study include: anti-p53 (#sc-126, Santa Cruz, 1:1000), anti-p21 (#sc-6246, Santa Cruz, 1:1000), anti-Nedd8 (#sc-373741, Santa Cruz, 1:1000), anti-GFP (#A02375-50, Genscript, 1:2000), anti-Phospho-p53 (Ser20) (#9287S, Cell Signaling Technology, 1:1000), anti-Phospho-p53 (Ser15) (#9284S, Cell Signaling Technology, 1:1000), anti-Acetyl-p53 (Lys379) (#2570S, Cell Signaling Technology, 1:1000), anti-MDM2(#51541S, Cell Signaling Technology, 1:1000), anti-ATM(#2873S, Cell Signaling Technology, 1:1000), anti-Phospho-ATM (Ser1981) (#13050S, Cell Signaling Technology, 1:1000), anti-CHK2(#3440S, Cell Signaling Technology, 1:1000), anti-Phospho-CHK2 (Thr68) (#2661S, Cell Signaling Technology, 1:1000), anti-BCL2(#15071S, Cell Signaling Technology, 1:1000), anti-Phospho-Histone H2A.X (Ser139) (#9718S, Cell Signaling Technology, 1:1000), anti-acetylated-lysine (#9441S, Cell Signaling Technology, 1:1000), anti-HA (#3724, Cell Signaling Technology, 1:1000), anti-β-actin (AC004, ABclonal, 1:2000), anti-α-Tubulin (AC012, ABclonal, 1:2000), anti-Flag (#SG4110-16, Shanghai Genomics Technology, 1:2000), anti-myc (SG4110-18, Shanghai Genomics Technology, 1:2000), anti-GAPDH (A19056, ABclonal, 1:2000), anti-Bax (A19684, ABclonal, 1:1000), anti-PUMA (A3752, ABclonal, 1:1000), anti-p53(10442-1-AP, Proteintech, 1:1000), anti-Sirt1(#07-131, Millipore,1:1000).

Techniques: Co-Immunoprecipitation Assay, Immunoprecipitation, Control, Western Blot, Transfection, Ubiquitin Proteomics, In Vitro, Purification, Incubation, Knock-Out, Activation Assay, Histone Deacetylase Assay, Activity Assay, Inhibition

Journal: Communications Biology

Article Title: The p53 R181C mutation accumulates through impaired deacetylation by Sirt1 and facilitates tumor development

doi: 10.1038/s42003-025-09465-y

Figure Lengend Snippet: A , B Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of genes with significant expression changes in both groups of samples (P < 0.05). C Statistics of differential peak distributions in samples from both groups. D – G Relative mRNA expression levels of PIK3CA , SHC1 , SRC, and PAK4 , in p53-KO HCT-116 transfected with Flag-vector, Flag-p53 WT or Flag-p53 R181C expression plasmids. Quantification data from three repeated experiments. Data are presented as mean ± SEM; *P < 0.05, ***P < 0.001, ****P < 0.0001 (Student’s t test). H – K ChIP-qPCR analyses of PIK3CA , SHC1 , SRC and PAK4 in p53-KO HCT-116 transfected with Flag-vector, Flag-p53 WT or Flag-p53 R181C. Quantification data from three repeated experiments. Data are presented as mean ± SEM; **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., no significance. (Student’s t test). L ChIP-seq profile of the PIK3CA , SHC1 , SRC , and PAK4 gene. M The Motifs of different peaks enriched, containing PIK3CA , SHC1 , SRC , and PAK4 gene.

Article Snippet: The antibodies used in this study include: anti-p53 (#sc-126, Santa Cruz, 1:1000), anti-p21 (#sc-6246, Santa Cruz, 1:1000), anti-Nedd8 (#sc-373741, Santa Cruz, 1:1000), anti-GFP (#A02375-50, Genscript, 1:2000), anti-Phospho-p53 (Ser20) (#9287S, Cell Signaling Technology, 1:1000), anti-Phospho-p53 (Ser15) (#9284S, Cell Signaling Technology, 1:1000), anti-Acetyl-p53 (Lys379) (#2570S, Cell Signaling Technology, 1:1000), anti-MDM2(#51541S, Cell Signaling Technology, 1:1000), anti-ATM(#2873S, Cell Signaling Technology, 1:1000), anti-Phospho-ATM (Ser1981) (#13050S, Cell Signaling Technology, 1:1000), anti-CHK2(#3440S, Cell Signaling Technology, 1:1000), anti-Phospho-CHK2 (Thr68) (#2661S, Cell Signaling Technology, 1:1000), anti-BCL2(#15071S, Cell Signaling Technology, 1:1000), anti-Phospho-Histone H2A.X (Ser139) (#9718S, Cell Signaling Technology, 1:1000), anti-acetylated-lysine (#9441S, Cell Signaling Technology, 1:1000), anti-HA (#3724, Cell Signaling Technology, 1:1000), anti-β-actin (AC004, ABclonal, 1:2000), anti-α-Tubulin (AC012, ABclonal, 1:2000), anti-Flag (#SG4110-16, Shanghai Genomics Technology, 1:2000), anti-myc (SG4110-18, Shanghai Genomics Technology, 1:2000), anti-GAPDH (A19056, ABclonal, 1:2000), anti-Bax (A19684, ABclonal, 1:1000), anti-PUMA (A3752, ABclonal, 1:1000), anti-p53(10442-1-AP, Proteintech, 1:1000), anti-Sirt1(#07-131, Millipore,1:1000).

Techniques: Expressing, Transfection, Plasmid Preparation, ChIP-qPCR, ChIP-sequencing

Journal: Communications Biology

Article Title: The p53 R181C mutation accumulates through impaired deacetylation by Sirt1 and facilitates tumor development

doi: 10.1038/s42003-025-09465-y

Figure Lengend Snippet: A mRNA was extracted from third-generation wild-type and p53R178C MEF cells, and real-time fluorescence quantitative PCR was used to measure the mRNA levels of downstream tumor suppressor genes p21, bax , and PUMA . B The expression level of p21 protein was measured in the heart, liver, spleen, lung, and kidney tissues of wild-type and p53R178C mice. C Expression levels of p53 protein in embryonic fibroblasts of wild type and p53R181C mutant mice during cell passaging. D The expression levels of p21 protein were measured in wild-type and p53R178C MEF cells during cell passaging. E Expression levels of p53 protein in third generation embryonic fibroblasts of wild type and p53R178C mutant mice under DNA stress conditions with or without exposure to IR. Extraction of mRNA from third generation embryonic fibroblasts of wild type and p53R178C mutant mice, and detection of p53 mRNA levels using real-time fluorescence quantitative PCR under DNA damage stress conditions with or without exposure to IR. F Third-generation wild-type and p53R178C MEF cells were tested for the proportion of cells entering S phase with and without IR. G Second-generation and sixth-generation wild-type and p53R178C MEF cells were stained with β-galactosidase to assess the degree of cellular senescence. Scale bar: 200px. H Third-generation wild-type and p53R178C MEF cells were tested for the expression of the apoptosis markers bax, Bcl-2 and PUMA proteins, as well as the senescence marker p21 protein, with and without IR. I Third-generation wild-type and p53R178C MEF cells were exposed to IR to assess the degree of cellular apoptosis. J Flag-p53WT and Flag-p53R181C were transfected into p53 knockout HCT116 cells, and tumor cell apoptosis was measured with or without DNA damage stress (DOX).

Article Snippet: The antibodies used in this study include: anti-p53 (#sc-126, Santa Cruz, 1:1000), anti-p21 (#sc-6246, Santa Cruz, 1:1000), anti-Nedd8 (#sc-373741, Santa Cruz, 1:1000), anti-GFP (#A02375-50, Genscript, 1:2000), anti-Phospho-p53 (Ser20) (#9287S, Cell Signaling Technology, 1:1000), anti-Phospho-p53 (Ser15) (#9284S, Cell Signaling Technology, 1:1000), anti-Acetyl-p53 (Lys379) (#2570S, Cell Signaling Technology, 1:1000), anti-MDM2(#51541S, Cell Signaling Technology, 1:1000), anti-ATM(#2873S, Cell Signaling Technology, 1:1000), anti-Phospho-ATM (Ser1981) (#13050S, Cell Signaling Technology, 1:1000), anti-CHK2(#3440S, Cell Signaling Technology, 1:1000), anti-Phospho-CHK2 (Thr68) (#2661S, Cell Signaling Technology, 1:1000), anti-BCL2(#15071S, Cell Signaling Technology, 1:1000), anti-Phospho-Histone H2A.X (Ser139) (#9718S, Cell Signaling Technology, 1:1000), anti-acetylated-lysine (#9441S, Cell Signaling Technology, 1:1000), anti-HA (#3724, Cell Signaling Technology, 1:1000), anti-β-actin (AC004, ABclonal, 1:2000), anti-α-Tubulin (AC012, ABclonal, 1:2000), anti-Flag (#SG4110-16, Shanghai Genomics Technology, 1:2000), anti-myc (SG4110-18, Shanghai Genomics Technology, 1:2000), anti-GAPDH (A19056, ABclonal, 1:2000), anti-Bax (A19684, ABclonal, 1:1000), anti-PUMA (A3752, ABclonal, 1:1000), anti-p53(10442-1-AP, Proteintech, 1:1000), anti-Sirt1(#07-131, Millipore,1:1000).

Techniques: Fluorescence, Real-time Polymerase Chain Reaction, Expressing, Mutagenesis, Passaging, Extraction, Staining, Marker, Transfection, Knock-Out