Fig. 2.

RhoC knockdown decreases LyEC proliferation and cyclin D expression in LyECs and HBMVECs, as well as increasing HBMVEC migration. (A) HBMVECs were transfected with control or RhoC siRNA (si) for 48 h, serum-starved overnight, and treated with VEGF-A (+V) for 16 h. Cyclin D1, RhoC and β-actin (loading control) were detected by western blotting of cell lysates. (B) 5×10⁴ serum-starved HUVECs treated with control or RhoC siRNA were seeded into collagen-coated Transwell chambers overnight and inserted into 24-well plates containing low-serum EGM. 10 ng/ml VEGF-A was added in the lower chamber and a Transwell migration assay was performed for 4 h. ***P<0.0001, paired two-tailed Student's t-test. (C,D) LyECs (C) or HBMVECs (D) were transfected with control or RhoC siRNA using Oligofectamine for 48 h. 4×10⁴ cells were plated in a 24-well plate, serum-starved (0.2%) overnight and treated with 10 ng/ml VEGF. Thymidine incorporation assays were performed. **P<0.05, paired two-tailed Student's t-test (RhoC siRNA −VEGF treated group versus control siRNA −VEGF treated group); *P<0.10 (RhoC +VEGF group versus control siRNA +VEGF group); NS, not significant. Results in B (n≥7) and C,D (n=2) are mean±s.d.