Fig. 6: STUB1-GOT2 axis promotes mitochondrial aspartate (Asp) synthesis and regulates mitochondrial dysfunction.

A The schematic showing the metabolic process of Asp. B The HEK293T cells transfected with the indicated plasmids for 48 h, then cell lysates were immunoprecipitated with anti-Myc magnetic beads, and immunoblotted with the indicated antibodies. C T24 or UC3 cells transfected with indicated plasmids for 48 h, Cell lysates were analyzed by western blotting using the indicated antibodies. T24 cells (D) or UC3 cells (E) were transfected with indicated plasmids for 48 h, Cell lysates were analyzed by Asp assay kit. T24 cells (F) or UC3 cells (G) were transfected with indicated plasmids for 48 h, Cell lysates were analyzed by ATP assay kit. H, I Immunofluorescent staining of reactive oxygen species (blue) with ROS Assay Kit in the T24 cells (J) or UC3 cells (K) transfected with indicated plasmids. J, K Immunofluorescent staining of mitochondrial membrane potential (red) with mitochondrial membrane potential assay Kit and nuclear (blue) with Nuclear 4’, 6-diamidino-2-phenylindole in the T24 cells (H) or UC3 cells (I) transfected with indicated plasmids. L, M T24 cells (F) or UC3 cells (G) were transfected with indicated plasmids for 48 h and then the cells were performed by colony formation assay. The p values were obtained by two-tailed unpaired t test. ^*p < 0.05, ^**p < 0.01.