Down-regulation of αPix levels attenuate hind-limb ischemia-induced Rac1 activation and angiogenesis. (A) Tissue extracts were prepared from ischemic and non-ischemic adductor muscles of WT and 12/15-Lox^−/− mice and analyzed for Src and αPix activation. To determine Src activation, its tyrosine phosphorylation was measured by Western blotting using its phospho-specific antibodies. Activation of αPix was measured as described in Figure 4A. (B) Tissue extracts were prepared from ischemic and non-ischemic adductor muscles of WT mice that received scrambled or αPix siRNA and analyzed for Rac1 activation by pull-down assay and αPix levels by Western blotting. (C) Blood perfusion was measured by LDPI in WT mice that received scrambled or αPix siRNA in response to ischemia. The bar graph represents the mean ± SD values of 3 independent experiments or 6 animals. *P < .05 versus WT non-ischemia; #P < .05 versus 12/15-Lox^−/− non-ischemia; **P < .05 versus WT ischemia; $P < .05 versus WT ischemia + scr siRNA.