FIGURE 2. DAPK forms two distinct heterocomplexes with HSP90 and CHIP or with DIP1/Mib1.
HEK293 cells were transfected with pCDNA3-human DAPK, pCDNA-His6-CHIP, or p3×FLAG-DIP. A, DAPK was immunoprecipitated (IP) with anti-DAPK antibody. B, CHIP was immunoprecipitated with anti-His tag antibody. C, DIP was immunoprecipitated with anti-DIP antibody. Western blots (WB) were probed with anti-DAPK, anti-HSP90, anti-CHIP, and anti-DIP1/Mib1 antibodies to detect recombinant human DAPK (r(hu)DAPK), recombinant CHIP (rCHIP), recombinant DIP1/Mib1 (rDIP), or endogenous HSP90. The blot is representative of three independent analyses. CE, cell extract; NS, nonspecific band.