Cell Culture Conditions for Breast Cancer Cell Lines

MDA-MB-231, MDA-MB-231/sh-ATG5 (ATG5 knockdown [5 (link)]), MDA-MB-231/GFP-LC3 (stable GFP-LC3 overexpression [5 (link)]) and MCF-7 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Cellgro, 10-013-CV) containing fetal bovine serum (10%; Gibco, 26140), penicillin (100 U/ml) and streptomycin (100 μg/ml) (Gibco, 15240) at 37°C and 5% CO₂ in a humidified incubator. T47D cells were cultured in RPMI 1640 medium under similar conditions. MCF-10A cells were maintained in DMEM/F12 media supplied with 5% horse serum, 20 ng/mL EGF, 0.5 mg/mL hydrocortisone, 100 ng/mL cholera toxin, 10 μg/mL insulin (Invitrogen, Carlsbad, CA, USA). Rapamycin (R8781), tamoxifen (T5648), bafilomycin A1 (B1793) and Chloroquine diphosphate salt (C6628) were from Sigma. Recombinant ADI-PEG20 was a gift of Polaris Pharmaceuticals Inc. (San Diego, CA).