SMCR7/MID49 Antibody - #DF12044

Figure 4 Sfrp5 decreased apoptosis and improved mitochondrial dysfunction. (A) Cardiomyocyte apoptosis determined by TUNEL staining. Scale bars = 25 μm. (B) Statistical analysis of TUNEL-positive cells (values are presented as mean ± SD, n = 6 per group). (C) Protein expression of Bax and Bcl-2 in mouse hearts at 14 days after MI. (D) Quantitative analysis of the relative Bcl-2/Bax ratio (n = 5 per group). (E) Macrophage infiltration determined by immunohistochemistry. Scale bars = 50 μm. (F) Statistical analysis of F4/80-positive cells (values are presented as mean ± SD, n = 6 per group). (G) IL1β mRNA expression (n = 6 per group). β-Actin was used as a loading control (values are presented as mean ± SD, n = 6 per group). (H) Quantitative analysis of the NADH oxidase activity level (values are presented as mean ± SD, n = 6 per group). (I) Quantitative analysis of the ATP level (values are presented as mean ± SD, n = 6 per group). (J) Quantitative analysis of the NAD+/NADH ratio (values are presented as mean ± SD, n = 5–6 per group). (K) Mitochondrial morphology was detected by TEM. Scale bars = 1 μm. (L) Quantitative analysis of the size of the mitochondria (values are presented as mean ± SD, n = 6 per group). (M) Quantitative analysis of the mitochondrial number/total area (values are presented as mean ± SD, n = 6 per group). (N) Protein expression of mitochondrial fusion proteins (MFN1 and MFN2) and mitochondrial fission proteins (p-Drp1Ser616, p-Drp1Ser616/Drp, Mid49, MFF, and Fis1) in mouse hearts at 14 days after MI (n = 5 per group). (O) Quantitative analysis of the relative MFN1 protein expression. Tubulin was used as a loading control. (P) Quantitative analysis of the relative MFN2 protein expression. Tubulin was used as a loading control. (Q) Quantitative analysis of the relative p-Drp1Ser616. Tubulin was used as a loading control. (R) Quantitative analysis of the relative p-Drp1Ser616/Drp. (S) Quantitative analysis of the relative Mid49 protein expression. Tubulin was used as a loading control. (T) Quantitative analysis of the relative MFF protein expression. Tubulin was used as a loading control. (U) Quantitative analysis of the relative Fis1 protein expression. Tubulin was used as a loading control. *p < 0.05 AAV9-NC-MI mice at 14 days after MI vs. AAV9-Sfrp5-MI mice at 14 days after MI. **p < 0.01 AAV9-NC-MI mice at 14 days after MI vs. AAV9-Sfrp5-MI mice at 14 days after MI. ***p < 0.001 AAV9-NC-MI mice at 14 days after MI vs. AAV9-Sfrp5-MI mice at 14 days after MI.

Fig. 5.| HIIT and MICT ameliorated mitochondrial fission and fusion in the hippocampus of APP/PS1 transgenic mice. The levels of DRP1 (a), FIS1 (b), MFF (c),MID49 (d), MID51 (e), MFN1 (f), MFN2 (g) and OPA1 (h) were detected in the hippocampus.

Figure 3 Mitochondrial changes in activated microglia (M1). (a) Schematic diagram of intracellular mitochondrial fusion and fission process in microglia after OGD/R. (b) Western blot assay of mitochondrial fusion protein (Opa1 and Mfn1), TOM20, and cytochrome c in mitochondria of M0 and M1 microglia (n = 3). Results are displayed in a form of mean ± SD; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. (c) Western blotting findings of mitochondrial fission protein including MFF, Fis1, Mid49, and Mid51 in mitochondria of M0 and M1 microglia (n = 3). Data presented are mean ± SD; ∗∗P < 0.01 and ∗∗∗P < 0.001. (d) Mitochondrial function in M1 microglia and M0 microglia was investigated by determining ATP (n = 3), mitochondria membrane potential (n = 3), and ROS (n = 3). The relative ratio of intracellular ATP was determined by calculating the ratio of level of ATP in M0-BV2 and M1-BV2 cells to level of ATP in M0-BV2 cells. Results are displayed in a form of mean ± SD. ∗∗P < 0.01 and ∗∗∗P < 0.001. (e) Morphology of intracellular mitochondria in BV2 cells visualized under TEM, scale bar: 1.0 μm.