Research Paper Volume 13, Issue 4 pp 6194—6204

USP30 protects against oxygen-glucose deprivation/reperfusion induced mitochondrial fragmentation and ubiquitination and degradation of MFN2

Effect of MFN2 overexpression on mitochondrial morphology in SK-N-BE(2) cells exposed to OGDR. SK-N-BE(2) cells were transfected with MFN2-Myc(200ng or 500ng) and Mito-GFP plasmids. After transfection 36 h, cells were treated with 4 h OGD plus reperfusion. (A) Digital photomicrograph under fluorescent illumination showed the morphology of mitochondria by mito-GFP. Most SK-N-BE(2) cells transfected with vector without OGDR displayed normal mitochondria, while fragmented mitochondria were evident in SK-N-BE(2) cells transfected with vector subjected to 4 h reperfusion after 4 h OGD. However, MFN2(200ng or 500ng) transfection significantly increased the number of SK-N-BE(2) cells with typical tubular and long mitochondria in a dose-dependent manner. (B) Quantitation (Mean ± SEM) of A from three independent experiments. Transfection with MFN2(200ng or 500ng) significantly attenuated OGDR-induced fragmentation of mitochondria in a dose-dependent manner. OE: over expression.

Figure 4. Effect of MFN2 overexpression on mitochondrial morphology in SK-N-BE(2) cells exposed to OGDR. SK-N-BE(2) cells were transfected with MFN2-Myc(200ng or 500ng) and Mito-GFP plasmids. After transfection 36 h, cells were treated with 4 h OGD plus reperfusion. (A) Digital photomicrograph under fluorescent illumination showed the morphology of mitochondria by mito-GFP. Most SK-N-BE(2) cells transfected with vector without OGDR displayed normal mitochondria, while fragmented mitochondria were evident in SK-N-BE(2) cells transfected with vector subjected to 4 h reperfusion after 4 h OGD. However, MFN2(200ng or 500ng) transfection significantly increased the number of SK-N-BE(2) cells with typical tubular and long mitochondria in a dose-dependent manner. (B) Quantitation (Mean ± SEM) of A from three independent experiments. Transfection with MFN2(200ng or 500ng) significantly attenuated OGDR-induced fragmentation of mitochondria in a dose-dependent manner. OE: over expression.