FIGURE 3.

Inhibition CaMKK β activity with STO-609 down-regulated SIRT1 phosphorylation under OGD and OGD with reoxygenation conditions in HBEC-5i endothelial cells. (a) and (b) Representative western blot images and quantitation of SIRT1 and phosphorylated SIRT1 (P-SIRT1, Ser-27) expression in HBEC-5i cells treated with DMSO (0.1%) or STO-609 (10 μM) under normoxia or 18-hr OGD. (c) and (d) Representative Western blot images and quantitated data analysis of SIRT1 and phosphorylated SIRT1 (P-SIRT1, Ser-27) expression in HBEC-5i cells treated with DMSO (0.1%) or STO-609 (10 μM) under normoxia or 18-hr OGD with 24 hr of reoxygenation (OGD18 hr + Reox24 hr). Quantitation of SIRT1 or P-SIRT1 was normalized to β-actin levels. Cells treated with DMSO and without OGD treatments were considered control cells (Ctrl). (e) and (f) Normalized pSIRT1 to total SIRT1 ratio under OGD18 hr or OGD18 hr + Reox24 hr. Data are expressed as the means ± SEM of at least three independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 versus DMSO-treated cells under normoxic conditions. ^#p < 0.05 and ^###p < 0.001 STO-609-treated cells under OGD or OGD with reoxygenation conditions. Statistical analyses were performed using one-way ANOVA and the Holm-Sidak multiple comparison test