Investigating mTOR and GSK3 Signaling

All mTOR kinase inhibitors, the GSK3 inhibitor SB216763, the proteasome inhibitor MG132, the protein synthesis inhibitor cycloheximide (CHX) and the antibodies against GSK3α/β, phospho-GSK3α/β (Ser21/9), phospho-Akt (Ser473), Akt, rictor and raptor were the same as described previously ^{22 (link)}. The GSK3 inhibitor CHIR99021 was purchased from LC laboratories (Woburn, MA). BKM120 was supplied by Novartis Pharmaceuticals Corporation (East Hanover, NJ). API-1 (NSC177233) was obtained from the National Cancer Institute (Bethesda, MD). MK2206 was purchased from Active Biochem (Maplewood, NJ). Perifosine was supplied by Keryx Biopharmaceuticals, Inc (New York, NY). Oil Red O was purchased from Sigma Chemical Co. (St. Louis, MO). SREBP1 (sc-13551), FASN (sc-55580) and α-tubulin (sc-23948) antibodies were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). ACC antibody (#3662) was purchased from Cell Signaling Technology, Inc. (Beverly, MA). Polyclonal Sin1 antibody (A300-910A) was purchased from Bethyl Laboratories, Inc. (Montgomery, TX).