Fig. 1. Experimental protocol outline. Cells were starved by serum reduction for 16h before pre-incubation with an SGLT2i (1 µM) or vehicle (0.02% DMSO) for 2h. Stimulation with 10 ng/ml TNFα occurred for 4-24h. Cells were treated for 4h for detection of adhesion molecules (ICAM-1, VCAM-1) and intracellular ROS levels, for 6h and 24h for western blot analysis, ROS and NO. Permeability and cell staining were assessed after 24h.