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. 2022 Jun 13;18(10):4026–4042. doi: 10.7150/ijbs.73493

Figure 5.

Figure 5

Effects of AKAP1 overexpression on mtDNA replication and mitochondrial function in HG-treated podocytes. (A) Representative Western blot and quantitative analysis of AKAP1 and TFAM in the NG, HG, AV-AKAP1 and HG+AV-AKAP1 groups. (n=3, *p < 0.05, **p < 0.01). (B) Quantitative PCR analysis of AKAP1 and TFAM in the four groups. (n=3, *p < 0.05, **p < 0.01, ***p < 0.001). (C-D) Double fluorescent labeling and quantitative analysis of TFAM in the four groups. (Scale bars, 50 µm. n=6, ***p < 0.001). (E) Quantitative PCR analysis of mtDNA including ND1, ND4L, cyt-b, COX I and ATPase6 in the four groups. (n=3, *p < 0.05, **p < 0.01). (F) EdU and MitoTracker red staining in podocytes of the four groups. (Scale bars, 50 µm). (G) MMP assay via JC-1 staining in the different groups. (Scale bars, 100 µm). (H-I) Semiquantitative analysis of mitochondrial ROS production in the different groups. (Scale bars, 100 µm. n=6, *p < 0.05, ***p < 0.001). (J) Semiquantitative analysis of ATP production in the different groups. (n=4, *p < 0.05). (K) Quantitative analysis of cell apoptosis by flow cytometry in the different groups. (n=3, ***p < 0.001).