Lipid droplet staining with Nile red

Cells were cultured with medium supplemented with oleate (0.2 mM, Sigma, Darmstadt, Germany) in the presence or absence of CD36 inhibitor, sulfo-N-succinimidyl oleate Na (SSO, 25 µM, Sigma) for indicated time. The cells were then washed with phosphate buffered saline (PBS) and stained with Nile red (1:5000, Invitrogen, Carlsbad, California, USA) for 30 min at 37°C. The quantification of Nile red content was measured by flow cytometry.