Deletion of <i>YAP</i> results in decreased HUVEC cell number.

<p>A. HUVEC cell number plotted against time after siRNA transfection. Untransfected and RISC-free siRNA were the control groups. Cells were counted on an Accuri C6 flow cytometer at 24 h intervals after transfection. There was a significant decrease in cell number in siYAP treated HUVECs compared to control HUVECs 48 h post-knockdown and at later time-points. Data are plotted as mean +/- standard error. N = 3 for each timepoint; *, p < 0.001. B. Quantification of BrdU+ HUVECs upon siYAP transfection. Cells were incubated with BrdU for 30 min and then stained with a BrdU antibody for visualization and quantification (plotted as the ratio of BrdU+ cells as a percent of total cells counted). Significantly fewer BrdU+ HUVECs were detected in siYAP-treated cells. Error bars indicate standard errors. P-values reflect differences compared to RISC-free control. *, p < 0.04; **, p < 0.007. C. Immunoblot (IB) assessing the levels of PCNA, CASPASE3, cleavage CASPASE3 in control and <i>YAP</i> knockdown (YAP-KD) cells. Protein samples were collected on d 1, 2 or 3 post-siRNA transfection and analyzed by western blot. No CASPASE3 cleavage was detected on d 2, 3 or 4, suggesting that <i>YAP</i> knockdown did not affect cell survival. See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117522#pone.0117522.s001" target="_blank">S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117522#pone.0117522.s002" target="_blank">S2</a> Figs.