Autophagy activation and SREBP‐1 induction contribute to fatty acid metabolic reprogramming by leptin in breast cancer cells

MCF-7, T47-D, and MDA-MB-231 cell lines were purchased from American Type Culture Collection (ATCC,
Rockville, MD, USA) and maintained in Dulbecco’smodified Eagle’s medium (DMEM) (MCF-7 cells) or
RPMI 1640 (T47D and MDA-MB-231 cells) containing10% FBS and 1% penicillin–streptomycin. Cells were
routinely cultured in an incubator at 37 °C under ahumidified atmosphere of 95% O2 and 5% CO2
 Measurement of total cellular ATP
Cellular ATP levels were measured using Luminescent
ATP Detection Assay Kit (Abcam) according to manufacturer’s instructions. Briefly, cells were seeded in 96-wellwhite plates at the density of 2 9 104cells per well. Afterovernight incubation, cells were treated with leptin and anappropriate agent as indicated. Cells were then lysed by
incubation with detergent for 5 min to release andstabilize ATP. ATP levels were determined by measurement of the luminescence formed by oxidation of thesubstrate D-luciferin in presence of luciferase using amicroplate reader (Fluostar Optima, BMG Labtech Inc,Ortenberg, Germany).

感谢韩国庆尚南道杨南大学药学院引用文献