Fig. 4

RESV stimulated autophagy and alleviated apoptosis through activating Notch1 signaling in CSE-treated HUVECs. N1ICD was overexpressed by LV-N1ICD transfection or suppressed by DAPT or siRNA in HUVECs as described in the Methods section. Notch1 mRNA expression was detected by qPCR (a). N1ICD protein, autophagy related proteins (LC3b-I/II, Beclin1, p62, mTOR) and apoptotic protein (cleaved caspase3) were detected by Western blot. β-Actin was used as loading control (c, k). Bar charts show the quantification of N1ICD, Beclin1, LC3B-II/LC3b-I, P62, mTOR and cleaved caspase3. Values are presented as means ± SD (b, d, e, f, g, h, l, m, n, o, p, q). Cell apoptosis was detected by flow cytometry (j). Statistical analysis was calculated (i). *P < 0.05, **P < 0.01 compared between the marked groups