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Figure 7 | Vascular Cell

Figure 7

From: γ-Actin plays a key role in endothelial cell motility and neovessel maintenance

Figure 7

Effect of γ-actin knockdown on ROCK signalling. (A) Representative photographs of HMEC-1 endothelial cells treated for 72 h with control (top) or γ-actin siRNA (bottom) and co-stained with phalloidin (left) and anti-paxillin antibody. The merged photographs (right) show phalloidin in red, paxillin in green and DNA (DAPI) in blue. Inset shows a magnified view of paxillin staining in the lamellipodial region. Scale bar, 20 μm. (B-C) Scatter dot plots showing the thickness of actin stress fibres and the size of paxillin-containing adhesions in HMEC-1 endothelial cells treated for 72 h with control (o) or γ-actin siRNA (∆). Bars, means of at least 15 individual cells. (D) Representative immunoblots of HMEC-1 cell lysates following treatment with control and γ-actin siRNA for 72 h. Membranes were probed with anti-γ-actin, anti-GAPDH (loading control) and anti-phospho-myosin light chain 2 antibodies. (E) Histogram showing the relative levels of phosphorylated myosin light chain 2 in HMEC-1 cells 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 control siRNA- versus γ-actin siRNA-transfected cells. **, p < 0.01; ***, p < 0.001.

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