Impaired Mitochondrial Fusion and Oxidative Phosphorylation Triggered by High Glucose Is Mediated by Tom22 in Endothelial Cells

Figure 8

Downregulation of Mfn1 decreases mitochondrial fusion and induces mitochondrial dysfunction. HUVECs were transfected with Mfn1 siRNA to knock down the expression of Mfn1. NC represents the negative control. (a) The downregulation of Mfn1 was confirmed by real-time PCR (). (b) The expression of the Mfn1 protein was detected by Western blotting and quantification analysis (). (c) Representative confocal microscopy images of Mfn1 knockdown HUVECs stained with MitoTracker® Deep Red. Scale bar: 10 μm. (d) Quantification of the form factor and aspect ratio of the mitochondrial networks (). (e) Representative images of mitochondria visualized by transmission electron microscopy. Mitochondria shortened in length in the Mfn1 knockdown group. Scale bar: 200 nm. (f) Total ATP level in HUVECs. (g) MMP was analyzed by JC1 staining and quantified based on fluorescence intensities (). (h) Apoptotic cells were detected by Annexin V/PI staining (). All data are presented as (, , and vs. NC).

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