Figure 2

γ-actin knockdown in vascular endothelial cells. (A) Histogram showing γ-actin relative gene expression following treatment with control (white) and γ-actin siRNA (black) for 24 h as assessed by quantitative RT-PCR. β2-microglobulin was used as housekeeping gene. Columns, means of at least three individual experiments; bars, SE. Statistics were calculated by comparing γ-actin relative expression in control and γ-actin siRNA-treated HMEC-1 cells; **, p < 0.01. (B) Representative immunoblots of HMEC-1 (left) and BMH29L (right) cell lysates following treatment with control and γ-actin siRNA for 72 h. Membranes were probed with anti-β-actin, anti-γ-actin and anti-GAPDH (loading control) antibodies. (C) Histogram showing the relative protein expression γ-actin as determined by densitometry after normalization with GAPDH, following treatment with control (white) and γ-actin siRNA (black) for 72 h. Columns, means of at least four individual experiments; bars, SE. Statistics were calculated by comparing γ-actin expression level in control and γ-actin siRNA-treated cells; ***, p < 0.001.