Figure 5.

Na,K-β cytoplasmic tail interacts with annexin II in a PI3-kinase dependent manner. (A) Ten or 20 μg of GST-βCD was incubated with 1 mg of MSV-β cell lysate and the annexin II pulled down by GST-βCD was determined by immunoblotting. (B) GST-annexin II (GST-DAII) was incubated with MSV-β cell lysate as described in Materials and Methods. The blots were then probed with anti-Na,K-β antibody to detect Na,K-β interaction with annexin II. (C) Na,K-β from MSV-β and MSV-βΔCD-GFP cell lysates was immunoprecipitated (IP) with anti-Na,K-β antibody and analyzed by immunoblotting (IB) for the presence of annexin II. (D) MSV-β cells were treated with DMSO or LY294002 and lysed. One milligram of total protein was used for immunoprecipitating Na,K-β. The amount of annexin II bound to Na,K-β was detected by immunoblotting. The graph represents the mean ± SD of three independent experiments.