Fig 1: ARIs prevent H2O2 induced senescence phenotype. Human keratinocytes were treated with 10 µM H2O2 or Veh (dH2O) and ARIs for 48 h. (A) SA-β-gal staining was performed to assess positive senescent cells. (B) Skin Cells stained with 5 µM DiHydroethidium (DHE) dye for 30 min to detect cytosolic superoxide and (C) cells stained with 5 µM MitoSOX dye for 10 min to detect mito superoxide. (D) qPCR analysis for confirming gene expression of senescent markers CDKN1A, TP53 and SERPINE1. (E) Gamma H2A.X Staining and (F) CellEvent™ Senescence Green Detection staining in human keratinocytes treated with Veh (dH2O), 400 µM H2O2 for 1 h followed by assessment of cells at day 10 with or without ARI AT-001. (G) ELISA to measure cytokine release in condition medium collected at day 10 and (H) qPCR analysis for confirming gene expression of senescent markers CDKN1A, TP53 and SERPINE1 CDKN2A and LMNB1. Data are presented as the mean ± SEM. n = 4 per group obtained from two consecutive experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test was implemented for comparison between groups. p* < 0.05 vs. H2O2. p# < 0.05 vs. Veh.