Kdr but not Flt1 mediates VEGFA-induced Src-PLD1-PKCγ-cPLA₂ activation in HRMVECs. HRMVECs transfected with control, Kdr, or Flt1 siRNA (100 nM) and quiesced were treated with and without VEGFA (40 ng/mL) for 30 minutes and cell extracts were prepared. An equal amount of protein from the control and each treatment group was analyzed for pSrc, pPLD1, pPKCγ, and pcPLA₂ levels using their phosphospecific antibodies. The blots were reprobed with anti-Src, anti-PLD1, anti-PKCγ, anti-cPLA₂, anti-Flt1, anti-Kdr, and anti-β-tubulin antibodies for normalization. The bar graph represents the quantitative analysis of 3 independent experiments. The values are presented as mean ± SD. *P < .01 vs control siRNA; ^†P < .05 vs control siRNA + VEGFA; ^§P < .01 vs control siRNA + VEGFA.