RAF1 controls arterial gene expression via ERK signaling. (A) Effect of MEK and PI3K inhibition on arterial gene expression. HUVECs transduced with control, WT RAF1 or RAF1S259A lentiviruses were treated with DMSO, MEK inhibitor U0126 (10 μM), or PI3K inhibitor LY294002 (10 μM) for 24 hours. DLL4, EFNB2, and HEY2 expression was assessed by qPCR. Data represent mean ± SEM of 3 independent experiments. (B) Immunoblot of Control, RAF1 WT and S259A lentivirus-infected HUVECs were treated with DMSO, MEK inhibitor U0126, or PI3K inhibitor LY294002. (C) DLL4 qPCR of HUVECs treated with DMSO, MEK inhibitor U0126, or PI3K inhibitor LY294002 at indicated doses. (D) qPCR of HUVECs infected with control, WT RAF1, S259A, S338/339A, Y340/341F, or S621A lentiviruses. (E) Immunoblot of HUVECs infected with indicated lentiviruses. (F) DLL4 qPCR of HUVECs infected with ad-lacz, ME, or ME-LA adenoviruses. *P < .05; **P < .01. mRNA, messenger RNA; NS, not significant.
