FIGURE 1.

GST pulldown and co-immunoprecipitation experiments confirm findings from the arrays. A–E, GST pulldown was done to confirm the interaction of PRRG4-cytosolic domain with: MAGI-1 (A); WWTR1/TAZ (B); YAP (C); NEDD4L (D); and NEDD4 (E). WWTR1/TAZ, MAGI-1, and YAP are FLAG-tagged, and anti-FLAG antibody was used for Western blot. Overexpressed proteins are written at the top of lanes. The cytosolic domain of PRRG4 interacts with endogenous NEDD4L (D) and NEDD4 (E) in GST pulldown experiments. F, co-immunoprecipitation experiments show that the cytosolic domain of PRRG4 interacts with WW domain-containing YAP, WWTR1, and MAGI-1b but not with SH3 domain-containing Grb2 or cool1/βPIX. Anti-FLAG antibody was used to immunoprecipitate the FLAG-tagged proteins, and anti-PRRG4 antibody was used for immunoblotting the pulled down PRRG4. Co-expressed cytosolic PRRG4 could be co-immunoprecipitated with MAGI, YAP, and TAZ from HEK293 cell lysates but not with Grb2 or cool1/βPIX.