Figure 3 : NAC/AAP protected mitochondria from H2O2-induced apoptosis.

From: Synergistic Protection of N-Acetylcysteine and Ascorbic Acid 2-Phosphate on Human Mesenchymal Stem cells Against Mitoptosis, Necroptosis and Apoptosis

Figure 3

(A) The MMP was measured using JC-1 fluorescence imaging in hADMSCs. The JC-1 monomer was represented by green fluorescence, the JC-1 aggregate image was represented by red fluorescence, and the merged images were the combined of the green and red images. Control cells showed strong aggregated red fluorescence indicative of normal membrane potential. Scale bar = 100 mm. (B) Changes of MMP in NAC or/and AAP-pretreated hADMCS by flow cytometry. Fluorescence intensity shifted from the higher level to the lower one indicates the loss of MMP. Mitochondria depolarization is indicated by an increase in the red fluorescence intensity ratio. Quantitative analysis of the green/red fluorescence shows that the NAC and/or AAP decreased green fluorescence, while NAC and/or AAP inhibited H2O2-mediated mitochondrial permeability. (C) The expression and localization of apoptosis related mitochondrial proteins. Expression of BAX and cytochrome c proteins in hADMSCs pretreated with NAC or/and AAP were assessed by western blot. COX-IV and a-tubulin were used as mitochondrial and cytosolic internal controls, respectively. Summary of normalized values of cytochrome c and BAX levels in the mitochondrial and cytosolic fractions of hADMSCs were shown at the right. (D) Immunostaining of BAX (left) and cytochrome c (right) using a respective FITC-conjugated antibodies (green) and MitoTracker (red) as the indicator of mitochondria in NAC or/and AAP-pretreated hADMSCs. Scale bar = 10 µm. * p < 0.05, ** p < 0.01, *** p < 0.001.