Fig. 4

Quantification of reduced glutathione (GSH) and ROS levels by DHE by immunofluorescence: a IF analysis using anti-GSH-NEM adduct-ab (1:500; v/v) showing decreased GSH levels in WT and Nrf2^−/− under sedentary and after HIES. Blue: nucleus (DAPI); Green: Anti-GSH staining. b Intensity analysis of the Anti-GSH staining. c Atrial tissue sections treated with cell-permeable DHE, the oxidized, fluorescent 2-hydroxyethidium (Red: DHE; Blue: DAPI). Images were obtained using confocal microscope at a magnification of 60x oil immersion. d ROS intensity was quantified using ImageJ Software and calculated as fold change of mean values of the WT sedentary group and the data is represented as mean ± SEM in the histograms (n = 3/group). *P < 0.05 vs SED WT; ^#P < 0.05 vs SED Nrf2^−/− and ^$P < 0.05 vs HIES WT