Fig. 7: Methylated UBE2m inhibits PPARγ-mediated fatty acid metabolism by neddylating NEDD4 in tubular epithelial cells.

A Representative western blot banding and quantitative analysis of the expression levels of NEDD4 from the indicated groups in HK-2 cells. B Detection of endogenous neddylation levels of NEDD4 from the indicated groups in mice or in HK-2 cells (C). D, E Detection of exogenous neddylation levels of NEDD4 in HEK193T cells with UBE2m knockdown or overexpression. F Detection of exogenous neddylation levels of NEDD4 in HEK193T cells transfecting UBE2m WT or RK. G Detection of endogenous neddylation levels of NEDD4 in UBE2mKO HK-2 cells re-introducing UBE2m WT or RK. H PLA detection of NEDD4 and NEDD8 interaction in HK-2 cells from the indicated groups. Scale bars = 50 μm. I HK-2 cells were treated with MLN4924. Representative western blot banding and quantitative analysis of the expression levels of UBE2m, NEDD4, and PPARγ from the indicated groups in HK-2 cells; and the expression levels of NEDD4, PPARγ, CD36, FATP2, CPT1α, and ACOX1 from the indicated groups in HK-2 cells (J). K Mitochondrial oxidative capacity was measured in real time after reintroduced UBE2m WT or RK treatment in UBE2m KO HK-2 cells, with or without MLN. L FAO blue staining demonstrates the degree of FAO in HK-2 cells from the indicated groups. Significance was assessed using two-way ANOVA tests. Data are shown as mean ± SD.