Anti rabbit 32460

Primary antibody sources: mCherry antibody (#43590), GFP antibody (#2956), FKBP1A/FKBP12 antibody (#55104), CD9 antibody (#13174), TSG101 antibody (#72312), and Alix antibodies (#18269) were diluted 1:1000 for western blotting, which was purchased from Cell Signaling Technology (Beverly, MA, USA); mTOR (human FRB Domain) antibody (diluted 1:1000 for WB, ALX-215-065) was procured from Enzo Life Science (Farmingdale, NY, USA); HSP70 antibody (diluted 1:1000 for WB, #242707) was obtained from R&D systems (Minneapolis, MN, USA); Beta actin antibody (diluted 1:1000 for WB, ab13772) was purchased from Abcam (Cambridge, MA, USA). Secondary antibody sources: anti-rabbit (#32460) and anti-mouse secondary antibody (#32430) were obtained from Thermo Fisher Scientific (Wilmington, DE, USA).

Rapamycin was dissolved in DMSO. The final concentration of DMSO in the culture medium did not exceed 0.2%. Anti-phospho-mTOR (ab109268, diluted 1:2,000) and anti-HIF-1α (ab51608, diluted 1:2,000) were from Abcam (Cambridge, UK). Anti-phospho-Akt (#4058, diluted 1:1,000) was from Cell Signaling Technology (Beverly, MA, USA) and anti-HSC70 (sc7298, diluted 1:10,000) was from Santa Cruz biotechnology (Dallas, TX, USA). Peroxidase-conjugated anti-mouse (#32430, diluted 1:2,000) and anti-rabbit (#32460, diluted 1:2,000) secondary antibody were from ThermoScientific (Waltham, MA, USA). Lysis buffer: [50 mM Hepes (pH 7.5), 150 mM sodium chloride, 1 mM EDTA (pH 8), 2.5 mM EGTA (pH 7.4), 0.1% Tween 20, 10% glycerol, 0.1 mM sodium orthovanadate, 1 mM sodium fluoride and 10 mM β-glycerophosphate] plus Protease inhibitor cocktail (#539134 Calbiochem, Darmstadt, Germany), PMSF and Phosphatase inhibitor Cocktail Set II (#524636 Calbiochem).