KLF2 reduces HUVEC migration without affecting VEGFR2-dependent VEGF signaling. The effect of a 7-day lentivirus-mediated overexpression of KLF2 on HUVEC migration was determined using a standard wounding assay. Untransduced (data not shown), Lenti-mock (○), and Lenti-KLF2 (•) transduced HUVEC cultures were wounded with a modified cell scraper, and photomicrographs were taken directly and after culturing the same cultures for an additional 24, 48, and 72 hours under standard conditions (A). Images were acquired using a 4 ×/0.13 NA objective. Migration distances were determined by measuring the width of the wounds at all time intervals (n = 3; B). (C) The effect of KLF2 on downstream VEGFR2 signaling was assessed by Western blotting using antibodies for total VEGFR2 and 2 specific phosphorylation states (Y951 and Y1054/1059). Following the 7 days after Lenti-KLF2 and Lentimock transduction, HUVEC cultures were treated with VEGF for 0, 2, and 24 hours before total protein lysates were made. Equal amounts of total proteins were loaded onto the gel and α-tubulin was used as an additional control for equal loading. *P < .01.