Fig. 3

iSGLT2 causes mitochondrion-mediated oxidative stress and apoptosis in CRC. Representative fluorescent images and cytofluorimetric detection of (A, B and H, I) intracellular and (C, D and J, K) mitochondrial ROS levels, and dot plots and analyses of (E, F and L, M) annexin V-FITC and propidium iodide (PI)-staining and (G, N) detection of caspase-3/7 activation in HCT 116 and HT-29 cells treated with 50 µM iSGLT2 for 72 h (iSGLT2). Control cells (Ctr) were maintained in complete culture medium with the corresponding volume of HBSS-10 mM Hepes. Scale bars = 100 µm. Lower left quadrant: viable cells; upper left quadrant: necrotic cells; lower right quadrant: early apoptotic cells; upper right quadrant: late apoptotic cells. *p < 0.05 vs. Ctr; †p < 0.01 vs. Ctr; ‡p < 0.001 vs. Ctr, by unpaired Student’s t-test