Fig. 2: Canagliflozin interferes with the ETC by inhibiting complex I.

a Oxygen consumption rate (OCR) and b extracellular acidification rate (ECAR) of a Seahorse Mito Stress Profile using differentiated RPTEC/TERT1 acutely exposed to the different gliflozins. Mean ± SD of a representative experiment performed in triplicate. c Key parameters of mitochondrial respiration. Mean ± SEM, n = 3 independent experiments performed in triplicate. *P < 0.05, ***P < 0.001 relative to 0.5% DMSO-treated assessed by one-way ANOVA with Bonferroni’s post-test. d Complex I-dependent respiration in permeabilized RPTEC/TERT1 cells exposed to 0.5% DMSO or 50 µM canagliflozin in the presence of either ADP (1 mM) alone or ADP with pyruvate (5 mM) and malate (2.5 mM), palmitoyl-L-carnitine (50 µM with 0.5 mM malate) or l-glutamine (4 mM with 0.5 mM malate). Mean ± SEM, n = 3 independent experiments. **P < 0.01, ***P < 0.001 relative to respective ADP only condition assessed by one-way ANOVA with Bonferroni’s post-test. e Complex II (10 mM succinate, 1 µM rotenone, 1 mM ADP) and f complex IV (1 µM rotenone, 1 µM antimycin A, 0.5 mM TMPD and 2 mM ascorbate) dependent respiration in permeabilized RPTEC/TERT1 cells exposed to 0.5% DMSO, 50 µM canagliflozin, 1 µM atpenin A5 or 10 mM NaN₃. Mean ± SEM, n = 3 independent experiments performed in duplicate. **P < 0.01, ***P < 0.001 relative to respective 0.5% DMSO-treated cells assessed by one-way ANOVA with Bonferroni’s post-test. g Complex I and h complex II activity in mitochondrial extracts exposed to the different gliflozins or the complex II inhibitor TTFA. Mean ± SEM, n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 relative to solvent control (s.c., 0.5% DMSO) assessed by one-way ANOVA with Dunnett’s post-test