FIGURE 1.
PKA is co-immunoprecipitated with WAVE2. A and B, FLAG-WAVE2 and HA-PKA Cα were transfected into COS-7 cells. The cell lysate was subjected to immunoprecipitation (IP) with anti-FLAG (A) or anti-HA (B) antibody. Coprecipitated proteins were visualized by immunoblotting using anti-FLAG and anti-HA antibodies. Vinculin was used as a loading control. C and D, endogenous WAVE2 was immunoprecipitated from MDA-MB-231 cells (C) or mouse brain extracts (D) with anti-WAVE2 antibody. Coprecipitated PKA was immunoblotted with anti-PKA C subunit antibody. E, cell lysate was isolated from COS-7 cells transfected with FLAG-WAVE2 and HA-PKA Cα. Following the addition of 8-bromo-cAMP (10−5 m), lysates were subjected to immunoprecipitation with anti-FLAG antibody. Proteins were visualized by immunoblotting using the indicated antibodies. F, the amount of coprecipitated PKA Cα subunit with WAVE2 (PKA Cα/WAVE2) was quantified by capturing film images and processing with ImageJ software (n = 3).