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. 2007 May 25;27(15):5296–5305. doi: 10.1128/MCB.01667-06

FIG. 1.

FIG. 1.

Identification and characterization of the interaction between Wwp2 and Rpb1. (A) Coomassie blue staining of an SDS-polyacrylamide gel electrophoresis gel containing the proteins from the indicated columns is shown. (B) GST pull-down assay to show that Wwp2 binds to Rpb1 directly in vitro. (C) Co-IP assay in HEK 293 cells to show that Wwp2 binds to the CTD of Rpb1 specifically in vivo. Whole-cell lysates (WCL) were immunoprecipitated (IP) with anti-Flag antibody and analyzed by Western blotting with antibodies as indicated. (D) Association of endogenous Wwp2 with Rpb1 in F9 cells. The RNAi Wwp2-2 F9 cells were induced with Tc for 3 days and then whole-cell lysates were subjected to immunoprecipitation with anti-Wwp2 antibody or control anti-GST antibody. (E) Association of endogenous Wwp2 with Rpb1 using F9 cell lysate treated with or without λ phosphatase (λ PPase). The whole-cell lysates of F9 cells were pretreated with λ PPase and subjected to IP as D.