Fig. 2

The sustained Ang II-induced formation of ROS in ECs is sensitive to a dual SGLT1 and SGLT2 inhibitor, sotagliflozin, and a selective SGLT2 inhibitor, empagliflozin. ECs are incubated with either (a, b) sotagliflozin (SOTA, 100 nM) or empagliflozin (EMPA, 100 nM) for 30 min before the addition of Ang II for either 30 min (a) or 24 h (b). For characterization of the role of SGLT1 and 2 in the pro-oxidant response to Ang II, ECs are incubated with Ang II for 24 h before being exposed to (c) the indicated glucose concentrations for 1 h in the presence or absence of sodium, (d) the indicated glucose concentrations, methyl α-D-glucopyranoside (AMG, a non-metabolizable glucose analogue), or mannitol, and (e) cariporide (a NHE-1 inhibitor, 10 µM), KB-R7943 (a NCX inhibitor, 10 µM), or ouabain (a NKA inhibitor, 10 nM) for 1 h, and the subsequent determination of dihydroethidium staining by confocal microscope. Results are shown as representative micrography of dihydroethidium staining (upper panels) and corresponding cumulative data (lower panels). Data are expressed as mean ± SEM of n = 3. ^*P < 0.05 vs. control and ^#P < 0.05 vs. Ang II