rVSMCs were stimulated with EGF (50 ng/ml), with or without 30 min pre-treatment of gefitinib (Gef, 1 µM), NS8593 (NS, 40 µM), 2-APB (30 µM) and ERK1/2 inhibitor PD98059 (PD, 20 µM). (A) Cell proliferation was assessed by CFSE proliferation assay using flow cytometry. Upper panel: Representative flow cytometry histograms. Lower panel: Corresponding scatter-plot graphs showing cell proliferation rate. Raw data were normalized to percentage differences based on the baseline value (veh). (B) Cell migration was examined by Scratch-wound assay. Cell migration was calculated as the percentage of wound healing closure at 20 h relative to 0 h time point. Upper panel: Representative images of rVSMCs migration stimulated with EGF in the presence and absence of different inhibitors. Lower panel: Corresponding scatter-plot graphs showing cell migration rate calculated as wound healing closure. Scale bar = 150 µm. *P<0.05 compared with vehicle control (veh, PBS) and ^†P<0.05 compared with EGF. Results represent the mean ± SEM of four to six independent experiments.