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. 2016 Mar 15;129(6):1234–1249. doi: 10.1242/jcs.179382

Fig. 7.

Fig. 7.

Downregulation of cofilin levels inhibit VEGFA-induced angiogenic events in HRMVECs. (A) Cells were transfected with control siRNA (siControl) or cofilin siRNA (siCofilin) and 36 h later cell extracts were prepared and analyzed by western blotting for cofilin and β-tubulin levels to show the efficacy of the siRNA on its target and off target molecules. (B–F) Cells were transfected with control or cofilin siRNA, growth-arrested and subjected to VEGFA (40 ng/ml)-induced F-actin stress fiber formation (B, scale bars: 50 μm), migration (C), DNA synthesis (D), sprouting (E) or tube formation (F) assays. (G) Cells were transduced with Ad-GFP or Ad-dnPak1 (40 MOI) or transfected with siControl or siPak1, allowed to become quiescent, and treated with and without VEGFA (40 ng/ml) for the indicated time periods. Then cell extracts were prepared. Equal amounts of protein from control and each treatment were analyzed by western blotting for phosphorylated (p)p70S6K and the blots were reprobed for p70S6K, Pak1 or β-tubulin levels to show either the overexpression, knockdown or normalization. The bar graphs represent quantitative analysis of three independent experiments. The values represent mean±s.d. *P<0.01 vs siControl; **P<0.01 vs siControl+VEGFA (one-way ANOVA followed by Tukey's post-hoc test).