BODIPY-FA uptake assay

HUVECS were seeded (1:1.9 density) on Ibidi arrays (1cm²) coated with 1% gelatin, followed by crosslinking with 0.5% glutaraldehyde, and grown to confluence with change of culture medium every other day. For stimulation, cells were incubated with 1% FF-BSA in plain M199 with inhibitors or vehicle, followed by stimulation with growth factors at indicated concentrations and intervals. The fluorescently labelled fatty acids (BODIPY-FA), C1-BODIPY^®500/510 C12 (20 μM) were added (3 minutes at 37°C) after washing the cells with 1% FF-BSA, PBS. Subsequently, the cells were fixed with 4% paraformaldehyde (PFA), as previously described [8]. They were imaged with 40x objective and equal exposure times.