histone h2ax [p ser139] antibody  (Bio-Techne corporation)

Journal: Advanced Science

Article Title: TPI1 Loss Triggers a Metabolite‐Driven Mitochondrial Redox Vulnerability via the SARM1–cADPR–Ca 2+ Axis

doi: 10.1002/advs.76614

Figure Lengend Snippet: TPI1 knockdown induces cellular senescence. (A) Volcano plot of transcriptomic alterations in TPI1 knockdown A498 cells. (B, C) GSEA analysis showing enriched pathways (B) and SASP signatures (C). (D) qRT‐PCR validation of senescence‐associated genes. (E) SA‐β‐gal staining in control and TPI1 knockdown cells with or without DHA treatment (1 mM, 72 h). (F) Immunoblot analysis of γH2AX, p53, p21, and LMNB1 in control and TPI1 knockdown cells. (G) Immunofluorescence analysis of indicated proteins in control and TPI1 knockdown A498 cells. (H) Cell cycle distribution was analyzed by flow cytometry in control and TPI1 knockdown cells. (I) Cytokine secretion measured by Luminex in control and TPI1 knockdown cells. (J) Cytokine secretion measured by ELISA in control and TPI1 knockdown cells. (K) Immunohistochemical analysis of γH2AX, p21, and IL‐6 in xenografts. (L) ROS levels measured by flow cytometry in control and TPI1 knockdown cells. (M) Immunoblot analysis of indicated proteins in TPI1 knockdown A498 cells with or without Mito‐TEMPO (5 µM, 24 h) treatment.

Article Snippet: After blocking with 5% skim milk, membranes were incubated with primary antibodies against Beta Actin (Proteintech, 66009‐1‐Ig), TPI1 (Proteintech, 10713‐1‐AP), TKFC (Proteintech, 12224‐1‐AP), LMNB1 (Proteintech, 66095‐1‐Ig), P21 (selleck, F0170), SARM1 (Proteintech, 28625‐1‐AP), CD38 (Zenbio, R381299), S6K1 (Proteintech, 14485‐1‐AP), Phospho‐S6K1 (Proteintech, 28735‐1‐AP), p53 (Proteintech, 60283‐2‐Ig), Phospho‐Histone H2AX (ABclonal, AP0687) at 4°C overnight.

Techniques: Knockdown, Quantitative RT-PCR, Biomarker Discovery, Staining, Control, Western Blot, Immunofluorescence, Flow Cytometry, Luminex, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining

Journal: Current Issues in Molecular Biology

Article Title: Nicotinamide Improves Skin Photoaging in Mice by Delaying Cellular Senescence and Suppressing the Senescence-Associated Secretory Phenotype

doi: 10.3390/cimb48070661

Figure Lengend Snippet: NAM inhibits the expression of cellular senescence biomarkers in photoaging mice. ( A – C ) The protein levels of γ-H2AX ( A ), p53 ( B ), and p21 ( C ) were determined by Western blot among UV, UV + 2% ( w / v ) NAM, UV + 5% ( w / v ) NAM, UV + 7.5% ( w / v ) NAM, and UV + 12.5% ( w / v ) NAM. ( D ) Quantification of the ratio of γ-H2AX, p53 and p21 to GAPDH. These results are means ± SD of three experiments. * p < 0.05, ** p < 0.01, *** p < 0.001. ( E ) Representative immunohistochemical images of p21. ( F ) The p21 positive cells were quantified. These results are means ± SD of three experiments. * p < 0.05, ** p < 0.01. p values ( D ) were calculated using a two-tailed unpaired t -test. p values ( F ) were calculated using one-way ANOVA followed by Tukey’s test in multiple groups.

Article Snippet: The primary antibodies used were anti-p21 antibody (ab188224, Abcam, Waltham, MA, USA), anti-p53 antibody (TA0865S, Abmart, Shanghai, China), anti-Collagen I antibody (ab255809, Abcam, Waltham, MA, USA), anti-ZO1 antibody (21773-1-AP, Proteintech, Wuhan, China), anti-E-cadherin antibody (20874-1-AP, Proteintech, Wuhan, China), anti-Phospho-Histone H2AX antibody (AP0687, ABclonal, Wuhan, China), anti-MMP12 antibody (A1709, ABclonal, Wuhan, China), anti-MMP1 antibody (10371-2-AP, Proteintech, Wuhan, China), anti-Phospho-TFEB antibody (87932S, CST, Danvers, MA, USA) and anti-GAPDH antibody (ab8245, Abcam, Waltham, MA, USA).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Two Tailed Test