Extended Data Fig. 6: Analysis of cellular respiration and activity of the respiratory complexes in GR-cKO versus control cardiomyocytes.
(a-c) Seahorse analysis of (a, n = 41 samples) basal respiration, (b, n = 6 samples) maximal respiration capacity and (c, n = 22 samples) extracellular acidification rate of cardiomyocyte cultures isolated from P1 control versus GR-cKO mice. (d) Specific activity of respiratory complexes (Complex I, CI; Complex II, CII; Complex III, CIII; Complex IV, CIV; Complex I+III, CI+III; Complex II+III, CII+III) normalized on citrate synthase (CS) activity, performed on mitochondria isolated from P7 GR-cKO and control hearts (n = 6 mice); (e-j) Seahorse analysis in response to (e-g) fatty acid oxidation inhibitor Etomoxir (Eto, 40 μM) or (h-j) inhibitor of glucose catabolism 2-deoxy-glucose (2dG, 5 μM) on P1 cardiomyocytes cultured in vitro (n = 7 samples in e, n = 8 samples in f, n = 8 samples in g; n = 7 samples in h; n = 7 samples in i; n = 7 samples in j); (k-m) Seahorse analysis of (k) acute response (n = 7 samples), (l) maximal respiration capacity (n = 7 samples) and (m) spare respiration capacity (n = 7 samples) in presence of 2-deoxy-glucose (2dG, 3 mM) in control versus GR-cKO P1 cardiomyocytes cultured in vitro. In all panels, numerical data are presented as mean (error bars show s.e.m.); statistical significance was determined using two-way ANOVA followed by Tukey’s test in h-j and two-sided Student’s t-test in l, m.
