AIP1 mediates TNF-α–induced ASK1 activation by facilitating dissociation of ASK1 from its inhibitor 14-3-3
J. Clin. Invest. Rong Zhang, et al. 111:1933 doi:10.1172/JCI17790 [Go to this article.]

Figure 5
Overexpression of AIP1 in ECs releases 14-3-3 from ASK1. (a) AIP1 dissociates 14-3-3 from ASK1. BAECs were transfected with expression plasmids for FLAG-tagged 14-3-3 (1 μg) with various amounts of FLAG–AIP1-N DNA (0, 0.1, 0.25, 0.5, 0.75, and 1 μg). Association of AIP1 and 14-3-3 with endogenous ASK1 was determined by immunoprecipitation with anti-ASK1 followed by Western blot with anti-FLAG. ASK1 in the immunoprecipitates was determined by Western blot with anti-ASK1. (b) AIP1-N does not bind to 14-3-3. Cell lysate expressing AIP1-N was used in a GST–14-3-3 pull-down assay. Bound AIP1-N was detected by Western blot with anti-FLAG. ASK1 was used as a control.