Fig. 6

SIRT6 is required for humanin to protect against HG-induced endothelial senescence. (A) Representative images of SA-β-gal staining and (B) quantification of SA-β-gal positive HUVECs treated with normal glucose (5.5 mM d-glucose, C), high glucose (15 mM d-glucose, HG), high glucose with [Gly14]-Humanin (15 mM d-glucose with 1 µM [Gly14]-Humanin, HG + HNG), normal glucose with OSS-128,167(5.5 mM d-glucose with 100 µM OSS-128167, C + OSS-128167), high glucose with OSS-128,167(15 mM d-glucose with 100 µM OSS-128167, HG + OSS-128167), high glucose with [Gly14]-Humanin with OSS-128,167 (15 mM d-glucose with 1 µM [Gly14]-Humanin with 100 µM OSS-128167, HG + HNG + OSS-128167). Scale bar = 20 μm. (C–E) The mRNA levels of (C) P53, (D) P16, (E) P21 of the six groups measured by qRT-PCR. (F) Representative images of Western Blotting analysis and (G-I) semi-quantification of (G) p53, (H) p21, and (I) Cyclin D1 of the six groups. (J) The representative images of wound healing assay of HUVECs of the six groups. (K) The semi‐quantitative analysis of the distance of migration. Scale bar = 100 μm. (L, M) Representative micrographs of EdU staining (L) and quantification (M) of EdU positive HUVECs. Scale bar = 100 μm. The values are expressed as the mean ± SEM. Experiments were carried out in triplicate and repeated three times. Results in B were analyzed with two-way ANOVA with Sidak’s multiple comparisons test. Results in C-E and K were analyzed with unpaired t test. Results in G-I and M were analyzed with two-way ANOVA with Dunnett’s multiple comparisons test.