Fig. 4.

CANA administration deteriorated cristae formation and reduced mitochondrial function. (a) MSCs were incubated with CANA (5 or 10 μM, 48 h) or DAPA (10 μM, 48 h). Intracellular ATP levels were determined via luciferin/luciferase-based assays (n = 3). (b) MSCs were incubated with CANA (10 μM, 48 h) or DAPA (10 μM, 48 h). Then, flow cytometry analysis was conducted to evaluate the fluorescence intensity of MitoSOX-stained MSCs (n = 3). The results are shown in a bar graph. (c) MSCs were incubated with CANA (10 μM, 48 h) or DAPA (10 μM, 48 h), and GDH1 activity was then evaluated using a spectrophotometric assay (n = 3). (d) MSCs were incubated with CANA (10 μM, 48 h). The oxygen consumption rates (OCRs) in each group of MSCs were assayed under both basal and maximal conditions (i.e., under CCCP treatment; n = 3). (e) MSCs were incubated with DAPA (10 μM, 48 h). The OCR of each group was detected as described in (d) (n = 3). (f) MSCs were incubated with CANA (10 μM, 48 h) or DAPA (10 μM, 48 h). Then, the MSCs were analysed by TEM. Representative images are shown (Scale bar=1 μm). Vacuole formation in cristae is indicated by a black arrow. (g) CANA- (10 μM, 48 h) or DAPA-treated (10 μM, 48 h) MSCs were stimulated with or without H₂O₂ (100 nmol) for 2 h. Cytochrome c release was then determined by Western blot analysis of both the cytosol (left) and mitochondrial pellets (right). VDAC and β-actin served as internal references. Data are means ± SDs, *P < 0.05; **P < 0.01, ***P < 0.001.