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Autophagy activation and SREBP‐1 induction contribute to fatty acid metabolic reprogramming by leptin in breast cancer cellsIF:6.60
34 人阅读发布时间：2026-01-30 16:46
Cellular ATP levels were measured using LuminescentATP Detection Assay Kit (Abcam) according to manu
facturer’s instructions. Briefly, cells were seeded in 96-wellwhite plates at the density of 2 9 104 cells per well. After
overnight 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 mea
surement of the luminescence formed by oxidation of thesubstrateD-luciferin in presence of luciferase using a
microplate reader (Fluostar Optima, BMG Labtech Inc,Ortenberg, Germany).
感謝韩国庆山市延南大学药学院 2 韩国庆山市延南大学细胞培养研究所分享文献！
Neutral lipid content was determined as previouslydescribed [33]. Cells were seeded at the density of
5 9 105 cells per 35-mm dish. After treatment withleptin and inhibitors as indicated, cells were incubated
with Bodipy 493/503 staining solution (2 µM) in darkat 37 °C for 20 min before a single cell suspension wascollected by trypsinization. Cells were then subjectedto flow cytometry analysis using BD FACSCaliburTM
(BD Biosciences, San Jose, CA, USA), and mean fluorescence intensity values were obtained by FLOWJO 7.6
Software (FlowJo LLC, Ashland, OR, USA).For visualization of intracellular lipid droplets, cells
were seeded in 8-well glass slides at the density of5 9 104 cells/well. After Bodipy staining as described
above, cells were fixed with 4% formaldehyde andcounterstained with DAPI. The images of lipid droplets were finally captured using a confocal microscopy(Nikon, Tokyo, Japan).
5 9 105 cells per 35-mm dish. After treatment withleptin and inhibitors as indicated, cells were incubated
with Bodipy 493/503 staining solution (2 µM) in darkat 37 °C for 20 min before a single cell suspension wascollected by trypsinization. Cells were then subjectedto flow cytometry analysis using BD FACSCaliburTM
(BD Biosciences, San Jose, CA, USA), and mean fluorescence intensity values were obtained by FLOWJO 7.6
Software (FlowJo LLC, Ashland, OR, USA).For visualization of intracellular lipid droplets, cells
were seeded in 8-well glass slides at the density of5 9 104 cells/well. After Bodipy staining as described
above, cells were fixed with 4% formaldehyde andcounterstained with DAPI. The images of lipid droplets were finally captured using a confocal microscopy(Nikon, Tokyo, Japan).