FIG 10.

Enhanced formation of Rac1/Pak2/MII complexes in CORO1A knockdown COS1 cells. (A to D) Anti-EGFP immunoprecipitates from parental (A to D) and CORO1 knockdown (C and D) COS1 cells transiently expressing the indicated combinations of proteins (A to D) and siRNAs (B) were analyzed by Western blotting to detect the amount of coimmunoprecipitated ectopic Rac1 and endogenous IQGAP1 under each experimental condition. (A and B) As a control, filters were stained with Ponceau solution prior to the immunoblotting step in some experiments to visualize the immunoprecipitated EGFPs. (C and D) Alternatively, the lysates were immunoblotted with antibodies to EGFP. Aliquots of the same cell lysates were analyzed by Western blotting to evaluate the expression of the indicated proteins prior to the immunoprecipitation step (bottom). Note that in panels C and D the increased amounts of immunoprecipitated IQGAP1 in lanes 3 and 5, respectively, are probably due to the slight variations in the amount of input lysate before the immunoprecipitation step (fifth gel from top).