Effect of CRIF1 deficiency on the mRNA levels of RAMPs and endogenous ADM2 in endothelial cells.

HUVECs were transfected with CRIF1 siRNA for 48 h. Next, the mRNA levels of (A) RAMP2 (B) RAMP3 and (C) ADM2 were determined by qPCR. (D) HUVECs were treated with the indicated doses of H₂O₂ for 24 h followed by the detection of ADM2 mRNA levels (E) HUVECs were pretreated with the antioxidants MitoTEMPO (1 nM) and NAC (5 mM) for 1 h followed by transfection with control or CRIF1 siRNA (100 pmol) for 48 h. mRNA levels of ADM2 were determined by qPCR. Data are means ± SD of three independent experiments. *P < 0.05, **P < 0.01 and *** P < 0.001 relative to the control, #P < 0.05 and ##P < 0.01 relative to siCRIF1 (n = 3 per group).