μ slide 6 0.5 glass bottom channel slides

EA.hy926 and HEK-293T cells were cultured in DMEM with 10% fetal bovine serum (FBS) and 1x penicillin/streptomycin (P/S) (referred to hereafter as complete DMEM). HUVECs were cultured using endothelial cell basal medium (EBM-2, Lonza) with the EGM-2 Bulletkit additives (Lonza) and were used for experiments between 0 and 5 passages. For serum starvation, EA.hy926 cells were cultured in DMEM with 0.5% FBS and 1x P/S. Cells were generally grown on plastic, tissue culture treated plates (Sarstedt) without any coating. For experiments, HUVECs or EA.hy926 cells were plated onto plastic dishes, glass-bottom 96-well plates, or μ-Slide VI 0.5 glass bottom channel slides (80607, ibidi) precoated with 5 μg/cm² of bovine fibronectin (MilliporeSigma) and 10 μg/cm² of rat tail collagen type I (MilliporeSigma). Plates were coated by incubation with proteins dissolved in sterile PBS for 1 h at 37 °C, followed by two rinses with sterile PBS, and were used immediately.