Research Paper Volume 10, Issue 7 pp 1666—1681

Figure 1. H₂S donor treatment is associated with partial rescue from cellular senescence phenotypes. Levels of the senescence-associated total CDKN2A gene expression (A) and levels its alternatively-expressed isoforms p14 and p16 (B) were assessed by qRTPCR in senescent endothelial cells after 24h treatment with H₂S donors (Na-GYY4137 at 100 µg/ml, AP39, AP123, RT01 at 10 ng/ml). Data are expressed relative to stable endogenous control genes GUSB, IDH3B and PPIA, and are given normalised to the levels of the individual transcripts as present in vehicle-only treated control cells. Fold change was calculated for in triplicate for three biological replicates. (C). The proportion of cells staining positive for Senescence Associated β-galactosidase (SA-β-Gal) activity following treatment with H₂S donors was determined by manually counting the percentage of SA-β gal positive cells. (D) The proportion of cells staining positive for H2Ax marker of DNA damage following treatment with H₂S donors was determined by manually counting the percentage of H2Ax positive cells. N = >300 cells for each sample. Statistical significance is indicated by *p<0.05, ** p<0.005, *** p<0.0001 (2 way ANOVA).