Figure 5

GILZ interferes with the mTORC2/AKT pathway. (a) Co-IP: M1-GILZ cells were lysed in CHAPS buffer, and immunoprecipitations (IP) were performed using anti-mTOR, anti-Rictor, anti-GILZ and control (nonspecific) antibodies. Immunoprecipitates and cell lysates were subjected to WB analysis using anti-mTOR, anti-Rictor, anti-HSP70 and anti-GILZ antibodies (b) As described in panel a, but with M1 cells pretreated with dexamethasone (DEX) for 24 h and using anti-Rictor, anti-GILZ and control IgG antibodies. (c) Same as in panel b, but using K562-r-GILZ cells. (d) Co-IP: M1 GILZ cells were lysed in CHAPS buffer, and immunoprecipitations were performed using anti-mSin1, anti-GILZ, anti-Rictor and control (nonspecific) antibodies. The immunoprecipitates and cell lysates were subjected to WB analysis using anti-Rictor, anti-mSin1 and anti-GILZ antibodies. (e) Same as in panel d, except that cells were lysed in a buffer that contained Triton X-100, which disrupts the interaction between mTOR and Rictor.