FIGURE 2.

Effect of IHPK2 mutation on binding of IHPK2 to TRAF2.a, NIH-OVCAR-3 cells were co-transfected with IHPK2 and TRAF2 by nucleofection. Pluses and minuses indicate the presence or absence of indicated construct, respectively. Total cell protein (100 μg) from each transfected cell line was immunoprecipitated with IHPK2 mouse monoclonal antibody. Precipitate was subjected to immunoblot analysis with TRAF2 polyclonal antibody (bottom row). Immunoblot analysis demonstrated that the IHPK2 S3&4 mutations in combination (but not single mutations) caused reduced TRAF2 binding when compared with the interaction of wild type IHPK2 and all other IHPK2 mutations. b, immunoblot to demonstrate protein levels of IHPK2, TRAF2, and GAPDH in lysates of transfected cells. The experiment was performed three times with similar results.