miR-34a inhibits endothelial cell proliferation, migration and tube formation.

<p><i>A:</i> miR-34a expression in endothelial cells expressing Bcl-2 (EC-Bcl-2) or vector alone (EC-VC) was analyzed by real time RT-PCR. <i>B-C:</i> Endothelial cells (EC) transfected with 50 nM of miR-34a or scrambled control RNA (SC) were used for different assays after 48 hrs of transfection. Endothelial cell proliferation <i>(B)</i> and cell migration <i>(C)</i> was measured using Xcelligence system. <i>D:</i> Endothelial cell tube formation assay was performed on Matrigel coated lab-tech chamber slides. At the end of assay, each chamber was photographed under microscope, and the area occupied by endothelial cell tubes was calculated using NIS-Elements-BS software (Nikon) and expressed as an angiogenic score <i>(E)</i>. Whole cell lysates were Western blotted (72 hrs post transfection) and probed with specific antibodies against E2F3a/b, SIRT1, survivin and CDK4 proteins. Equal protein loading in Western blotting was verified by stripping the blots and reprobing with tubulin antibody.