Mopc 21

INS-1 cells carrying the Tet-on system for CD36 overexpression (15 (link)) were cultured with RPMI 1640 medium containing 11.1 mmol/L glucose, 10% FBS, 100 IU/mL penicillin, 100 μg/mL streptomycin, 50 mg/mL neomycin, 50 mg/mL hygromycin, 10 mmol/L HEPES, 1 mmol/L sodium pyruvate, and 50 μmol/L β-mercaptoethanol at 37°C in a humidified atmosphere with 5% CO₂. To induce CD36 expression, cells were seeded in 24- or 48-well plates and cultured with or without 500 ng/mL doxycycline (Sigma-Aldrich, St. Louis, MO) for 72 h.
EndoC-βH1 cells (16 (link)) were cultured in Matrigel/fibronectin-coated (100 μg/mL and 2 μg/mL, respectively) (Sigma-Aldrich) vessels with DMEM containing 5.6 mmol/L glucose, 2% BSA, 10 mmol/L nicotinamide, 50 μmol/L β-mercaptoethanol, 5.5 μg/mL transferrin, 6.7 ng/mL sodium selenite, 100 IU/mL penicillin, and 100 μg/mL streptomycin at 37°C in a humidified atmosphere with 5% CO₂. Cells were seeded in Matrigel/fibronectin-coated 48-well plates and cultured for 72 h with 2 μg/mL of a CD36-blocking antibody (FA6.125, catalog number 60084; STEMCELL Technologies, Vancouver, British Columbia, Canada) or an isotype control (MOPC-21, catalog number ab18443; Abcam, Cambridge, U.K.).