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

Neutral lipid content was determined as previously
described [33]. Cells were seeded at the density of
5 9 105 cells per 35-mm dish. After treatment with
leptin and inhibitors as indicated, cells were incubated
with Bodipy 493/503 staining solution (2 µM) in dark
at 37 °C for 20 min before a single cell suspension was
collected by trypsinization. Cells were then subjected
to 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 of
5 9 104 cells/well. After Bodipy staining as described
above, cells were fixed with 4% formaldehyde and
counterstained with DAPI. The images of lipid droplets were finally captured using a confocal microscopy
(Nikon, Tokyo, Japan).

感谢韩国庆山市，延南大学细胞培养研究所引用文献