RAF1^S259A induces SEMA6A-dependent EC repulsion. (A) Repulsion assay of HUVECs infected with ad-GFP, RAF1 WT, and S259A. Green cells are HUVECs infected with ad-GFP, RAF1 WT, and S259A and transfected with indicated small interfering RNAs (siRNAs). Red cells are HUVEC infected with mCherry lentivirus. Colocalized areas were indicated as white. (B) Quantification of repulsion assay shown in (A). Data represent mean ± SEM of 3 independent experiments. (C) Repulsion assay of HUVECs after knocking down SEMA6A, NTNG1, and EFNB2. (D) Quantification of repulsion assay shown in (C). Data represent mean ± SEM of 3 independent experiments. Knocking down efficiency is shown in (E). mRNA, messenger RNA.