Fig. 4

iSGLT2 impairs energetic state and modulates SIRT3 levels in CRC. Evaluation of (A) GSH/GSSG ratio and (B) ATP production coupled respiration, (C) maximal and (D) basal respiration, and (E) coupling efficiency assessed by Seahorse analyzer and (F) SIRT3 activity, (G) content evaluated by ELISA kit, (H) mRNA expression and (I) immunoblotting analysis with cropped blots in HCT 116 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. M = molecular weight markers, lane 1 = Ctr, lane 2 = iSGLT2. Western blotting is expressed as arbitrary units (AU), mRNA levels are reported as floating bars with a line representing the median ± SD of n = 3 independent experiments. †p < 0.01 vs. Ctr; ‡p < 0.001 vs. Ctr, by unpaired Student’s t-test