Interaction of TPPP3 with VDAC1 Promotes Endothelial Injury through Activation of Reactive Oxygen Species

Figure 3

TPPP3 binds and stabilizes VDAC1. (a) Immunoprecipitation (IP) of cell lysates from the HUVECs transfected with TPPP3-FLAG (left) or VDAC1-HA (right) with antibodies to FLAG or HA, and Western blot analysis with the indicated antibodies to TPPP3 or VDAC1. Irrelevant rabbit IgG was used as control. (b) Representative confocal images of TPPP3 (green) and VDAC1 (red) are shown; colocalization of TPPP3 and VDAC1 was observed (yellow) in HUVECs. (c) HUVECs were transduced with vector or TPPP3 and then treated with CHX (10 μg/mL) for the indicated time. The expressions of VDAC1 and GAPDH were determined using Western blotting. (d) HUVECs were treated with 200 μM PA and then treated with CHX (10 μg/mL) for the indicated time. The expressions of VDAC1 and GAPDH were determined using Western blotting. (e) Western blot analysis of TPPP3, VDAC1, BCL-2, Bax, Cyto C, Caspase-3, and Caspase-9 proteins following overexpression of TPPP3 in HUVECs with or without 50 μM PA treatment. One-way ANOVA was performed to assess the difference. .

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