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 USP30 overexpression on the ubiquitination and protein expression of MFN2 in SK-N-BE(2) cells exposed to OGDR. SK-N-BE(2) cells were transfected with indicated plasmid. After transfection 36 h, SK-N-BE(2) cells were treated with 4 h OGD plus 4 h reperfusion. (A) After immunoprecipitation of MFN2, western blot was performed to examine the ubiquitination and protein level of MFN2 after 4 h reperfusion following 4 h OGD in SK-N-BE(2) cells. Transfection with USP30-Flag significantly attenuated OGDR induced ubiquitination of MFN2 and increased protein expression of MFN2. (B) Quantitation (Mean ± SEM) of ubiquitination of MFN2 from three independent experiments. (C) Quantitation (Mean ± SEM) of protein expression of MFN2 from three independent experiments. OE: over expression.

Figure 6. Effect of USP30 overexpression on the ubiquitination and protein expression of MFN2 in SK-N-BE(2) cells exposed to OGDR. SK-N-BE(2) cells were transfected with indicated plasmid. After transfection 36 h, SK-N-BE(2) cells were treated with 4 h OGD plus 4 h reperfusion. (A) After immunoprecipitation of MFN2, western blot was performed to examine the ubiquitination and protein level of MFN2 after 4 h reperfusion following 4 h OGD in SK-N-BE(2) cells. Transfection with USP30-Flag significantly attenuated OGDR induced ubiquitination of MFN2 and increased protein expression of MFN2. (B) Quantitation (Mean ± SEM) of ubiquitination of MFN2 from three independent experiments. (C) Quantitation (Mean ± SEM) of protein expression of MFN2 from three independent experiments. OE: over expression.