Figure 5. PAK4 overexpression decreases TNF-induced EMPs release. (A) Western blotting of HUVECs transfected 48 h with GFP, GFP-PAK4 plasmid, siPAK4 or GFP-PAK4+siPAK4, and analyzed by PAK4 and control tubulin antibodies. (B) EMP release in HUVECs transfected with GFP, GFP-PAK4, GFP-PAK4KD, siPAK4 or GFP-PAK4+siPAK4, and stimulated 24 h with TNF (100 ng/ml) or control vehicle. Overexpression of GFP-PAK4 significantly inhibits TNF-induced EMP release (lane 5 vs lane 3). The TNF-induced EMP is also inhibited by caspase inhibitor z-VAD (lane 7). PAK4 kinase mutant (GFP-PAK4KM) does not reduce the TNF-induced EMP release (lane 8). The values represent the mean of three independent experiments. ** P<0.01; NS, not significant.