Anti pan akt

Primary antibodies used were anti-phospho-Akt (Ser473; #4060, Cell Signaling), anti-pan-Akt (#2920, Cell Signaling), anti-phospho-P44/42 MAP(ERK1/2) (#9101, Cell Signaling), anti- P44/42 MAPK (ERK1/2) (L34F12) (#4696, Cell Signaling), anti-phospho-mTOR (SER2448 (D9C2), #5536, Cell Signaling), anti-mTOR (# 2972, Cell Signaling), anti-oxidative phosphorylation (OXPHOS) complexes I to V (#ab110413, Abcam), anti-KLF10 (#ab73537, Abcam), and β-actin (#4970, Cell Signaling).

The ECC-1 and Ishikawa cell lines were provided as a gift from Dr Bruce Lessey (Department of OB/GYN Greenville Memorial Hospital) (15 ). Both cell lines are estrogen receptor-alpha positive and progesterone receptor weakly positive, which was recently confirmed in our laboratory by chloramphenicol acetyltransferase (CAT) activity. The ECC-1 cells were maintained in RPMI 1640 containing 5% fetal bovine serum, 300 mM l-glutamine, 5 μg/ml bovine insulin, 10,000 U/ml penicillin and 10,000 μg/ml streptomycin under 5% CO₂. The Ishikawa cells were grown in MEM supplemented with 5% fetal bovine serum, 300 mM l-glutamine, 10,000 U/ml penicillin and 10,000 μg/ml streptomycin under 5% CO₂. Simvastatin, MTT (3-5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and RNase A were purchased from Sigma (St. Louis, MO). The anti-phosphorylated-AKT, anti-pan-AKT, anti-phosphorylated-p42/44, anti-pan-p42/44, anti-phosphorylated-S6, anti-pan-S6, anti-cleaved caspase 3, anti-BCL-2, and anti-MCL-1 antibodies were purchased from Cell Signaling (Beverly, MA). The anti-HMGCoA antibody was from Santa Cruz (Dallas, Texas). Enhanced chemiluminescence Western blotting detection reagents were purchased from Amersham (Arlington Heights, IL). All other chemicals were purchased from Sigma.