Fig. 1
Anti-oxidative effects of NAC. a(1) Cell viability graphs following NAC treatment at concentrations of 10, 100, 500, 1000 μM, and 10 mM for 24 h. All data are expressed as the mean ± SEM (N = 4). The dash-dotted line is a 10 mM NAC concentration producing 30% cell viability. a(2) This graph is an enlargement of a(1) below 1000 μM. The concentration of NAC showing less than 10% cytotoxicity was 1000 μM. a(3) The cell viability assay of H2O2. H2O2 more than 100 μM was cytotoxic, killing 60% of cells in the sample. b(1), c(1) An illustrative Western blotting picture of Nrf2 and phospho-p62 (p-p62) following NAC treatment (1000 μM) for 3 or 24 h. b(2), c(2) Normalized values of Nrf2 or p-p62 were graphed (N = 3). Normalization was performed with Nrf2 or p-p62 band intensity of untreated cell (3-h incubation). d Immunocytochemistry of MH7A cells stained to identify Nrf2. The color analyzed by confocal laser scanning microscopy is expressed as Nrf2 (green)/phalloidin (red)/DAPI (blue). d(1) Untreated cells were cultured for 24 h. d(2) Cells cultured for 24 h treated with low-dose NAC (1000 μM). Scale bar; 20 μm. e ROS formation in MH7A cells following H2O2 treatment (100 μM) for 1 h and low-dose NAC treatment (1000 μM) for 3 (e(1)) and 24 h (e(2)). All values are expressed as the mean ± SEM (N = 3). The significance levels are shown as *P value < .05, **P value < .01, ***P value < .001, ****P value < .0001