Temsirolimus cci 779

The antibodies used in this study include the following: rabbit anti-phosphor-mTOR, rabbit anti-mTOR, mouse anti-Phospho-p70 S6 Kinase, rabbit anti-p70 S6 kinase, rabbit anti-CD44, rabbit anti-cleaved PARP and rabbit anti-cleaved caspase-3 were all obtained from Cell Signaling Technology (Danvers, MA, USA). Mouse-anti-p32 (gC1qR) antibody was obtained from Abcam (Cambridge, MA, USA). Rabbit anti-Akt and mouse anti-Phospho-Akt antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat anti-mouse and anti-rabbit IgG-horseradish peroxidase-conjugates were from Pierce (Rockford, IL, USA). Mouse monoclonal anti-actin was a gift from Dr. José Manuel Hernandez-Hernández (CINVESTAV-IPN). Temsirolimus (CCI-779) and 5-fluorouracil (5-FU) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Hydroxychloroquine, temsirolimus (CCI-779), and 5-fluorouracil (5-FU) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The HIF-2α-specific inhibitor PT-2385 was purchased from Biovision. Dihydroethidium (DHE) was obtained from Invitrogen^TM (Waltham, MA, USA) (Cat. No. D11347). N-Acetyl-L-cystein (NAC) was obtained from Sigma-Aldrich (Cat. No. A7250). The antibodies used in the experiments were from the following sources: anti-LC3 from Novus Biologicals (Centennial, CO, USA); and anti-activated caspase-3, anti-HIF-1α, anti-HIF-2α, anti-SQSTM1/p62, anti-β-tubulin, anti-Atg7, anti-Phospho-S6 Ribosomal protein (Ser235/236), anti-Beclin.1 and anti-PI3K-Class III were all obtained from Cell Signaling Technology (Danvers, MA, USA). Goat anti-mouse and anti-rabbit IgG-horseradish peroxidase-conjugates were from Pierce (Rockford, IL, USA)