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. 2018 Sep 15;19(9):2782. doi: 10.3390/ijms19092782

Figure 3.

Figure 3

Refreshment of mitochondria in H9C2 cells and SS31 cyto-protective effect abolished by SIRT3 suppression. (A) Activity (OCR) of isolated mitochondria (determined by the Mito stress test kit and the XFe24 Analyzer) (n = 4). (B) Immunofluorescent (IF) (400×) staining showing nuclear DAPI stain (B-1,B-5), endogenous mitochondria (B-2,B-6), exogenous mitochondria (red color) being transferred into H9C2 cells (B-7), and (B-3) indicating that no exogenous mitochondria are transfused into H9C2 cells. (B-4) indicating merged of (B-1–B-3), and (B-8) indicating merged pictures of (B-5–B-7) with a yellow-green color indicating the fusion of exogenous and endogenous mitochondria in H9C2 cells. (C) Protein expression of NOX-1, * vs. other groups with different symbols (†, ‡), p < 0.0001. (D) Protein expression of NOX-2 * vs. other groups with different symbols (†, ‡), p < 0.0001. (E) Protein expression of mitochondrial Bax * vs. other groups with different symbols (†, ‡), p < 0.0001. (F) Protein expression cytosolic cytochrome C (cyt-Cyto C) * vs. other groups with different symbols (†, ‡), p < 0.0001. (G) Protein expression of mitochondrial cytochrome C (mit-Cyto C) * vs. other groups with different symbols (†, ‡), p < 0.0001. (H–K) Illustrating the microscopic finding (400×) of TUNEL assay for the identification of apoptotic nuclei (green color) in H9C2 cells. (L) Analytical results of the number of apoptotic nuclei * vs. other groups with different symbols (†, ‡), p < 0.0001. The blue color indicated nuclei stained by DAPI. All statistical analyses were performed by one-way ANOVA, which was followed by a Bonferroni multiple comparison post hoc test (n = 6 for each group). Symbols (*, †, ‡) indicate significance at the 0.05 level. Mena = menadione.