Fig. 4

Ang II-induced endothelial senescence is dependent on the AT1R/NADPH oxidase/SGLT1 and 2 pathways. ECs are incubated with either VAS-2870 (VAS, 1 µM), losartan (LOS, an AT1R antagonist, 1 µM), sotagliflozin (SOTA, 100 nM), empagliflozin (EMPA, 100 nM), cariporide (10 µM) or KB-R7943 (10 µM) for 30 min before the addition of Ang II, and the subsequent determination of (a) SA-β-gal activity, (b–d) the expression level of p53, p21 and p16 protein after a 24-h incubation period by Western blot analysis, and (e, f) the expression level of p53 and p21 mRNA after a 6-h incubation period by RT-PCR. Results are shown as representative immunoblots (upper panels) and corresponding cumulative data (lower panels). Data are expressed as mean ± SEM of n = 3. ^*P < 0.05 vs. control and ^#P < 0.05 vs. Ang II