Figure 5

Biochemical validation of potential human ING protein interactions. A) HEK293 cells were transfected with the equal amounts of the indicated constructs and cell lysates were immunoprecipitated using anti-ING1 and then immunoblotted with anti-p38MAPK. The blot was reprobed with anti-ING1 to confirm equal IP efficiency. B) Same as in panel A but immunoblotting with anti-MEKK4. C) Endogenous interaction between ING1 and p38 MAPK under non-UV and UV conditions. HEK293 cell lysates were immunoprecipitated using anti-ING and then immunoblotted with anti-p38MAPK. Interestingly, the interaction seems to be stronger under normal rather than stress conditions. The input shows equal amounts of cell lysates have been used. ING1 levels after reblotting of the same membrane with anti-ING1. ING1 levels were equal under both normal and stress conditions. D) Same as in C but the immunoblotting was done with anti-MEKK4 to detect ING1-MEKK4 interaction at the endogenous interaction. E) Same as in C but the immunoblotting was done with anti-RAD50 to detect ING1-RAD50 interaction at the endogenous level.