Effect of exogenous ADM2 on the Akt-CREB signaling pathway in CRIF1-silenced endothelial cells.

HUVECs were transfected with control or CRIF1 siRNA (100 pmol) followed by treatment with ADM2 for 48 h. Next, the mRNA levels of (A) RAMP2 (B) RAMP3 and (C) ADM2 were determined by qPCR. (D) Western blot analysis of phospho-Akt and phospho-CREB levels. (E and F) Protein levels were quantified by densitometric analysis using ImageJ software. Data are means ± SD of three independent experiments. *P < 0.05 and *** P < 0.001 relative to the control, #P < 0.05, ##P < 0.01 and ns = not significant relative to siCRIF1 (n = 3 per group).