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. 2012 Oct 31;7(10):e46889. doi: 10.1371/journal.pone.0046889

Figure 3. HSG modulates EC proliferation through the Src/Vav2 pathway.

Figure 3

(A) ECs were seeded on HSG and LSG for 24 h, and cell lysates were immunoblotted with antibodies against phospho-Src (Y416), Src, phospho-Vav2 (Y172), and Vav2. (B) ECs pretreated with DMSO and PP1 (20 µM) for 1 h were seeded on HSG for 1 h, 4 h or 24 h. Cell lysates were analyzed for phospho-Src (Y416), Src, phospho-Vav2 (Y172), and Vav2. (C) Vav2 was knocked down in ECs by shRNA (sh-Vav2) with more than 50% reduction in Vav2 RNA and protein levels (left lower and upper panels, respectively). Cells with sh-Vav2 and control-shRNA were seeded on HSG and LSG for 24 h, and cell lysates were subjected to an RBD-pull down assay with an antibody against RhoA (right panel). (D–E) ECs were pretreated with PP1 or transfected with Vav2-shRNA (sh-Vav2), seeded on HSG and LSG for 24 h, and subjected to immunoblotting for protein expression (D) or flow cytometry for cell cycle analyses (E). * p<0.05 in comparison with the corresponding controls on HSG and LSG. § p<0.05 for comparison between DMSO and PP1 or between control and sh-Vav2 on HSG (n = 3).