Fig. 4.

RhoC promotes proliferation through cell cycle progression. HUVECs were transfected with control or RhoC siRNA for 48 h, serum-starved overnight, and treated with VEGF-A for 16 h (A,B; +V16h) or 6 or 12 h (C, +V6h and +V12h, respectively). Experiments were repeated at least three times. (A) The cells were fixed, stained with propidium iodide, and analyzed by FACS. The mean±s.d. percentage of cells with DNA content in each of the three phases of the cell cycle is shown over three independent determinations. *P≤0.05, and **P≤0.001 (paired two-tailed Student's t-test). (B,D) Cyclin D1, cyclin A, cyclin B1, p27, p21, RhoC and β-actin (loading control) were detected by immunoblotting (IB) of cell lysates (B). Vertical lines indicate where lanes were removed and composite images were generated from the same immunoblot. Densitometry of the indicated immunoblots was performed using ImageJ software (D). (C) Nuclear fractions were collected and subjected to western blotting using an anti-β-catenin antibody.