Fig. 5: GLUL reduces the K48 ubiquitin connectivity level of TRM25 to UAP1.
A Representative Western blotting images of HEK-293T cells cotransfected with the TRIM25 overexpression vector and UAP1 in a dose-dependent manner. B Representative image of a Western blotting experiment showing UAP1 expression levels after cell transfection with si-TRIM25. C Representative image of a Western blotting experiment showing UAP1 expression levels after cell transfection with si-TRIM25. D Representative images of Western blotting analysis of proteins that were extracted from HEK293T cells that were transfected with the Myc-UAP1 and Flag-TRIM25 overexpression plasmids and treated with MG132. E Representative images of Western blotting analysis of proteins that were HEK293T cells that were transfected with the Myc-UAP1 and Flag-TRIM25 overexpression plasmids and treated with MG132 and chloroquine. F A Myc-UAP1 overexpression plasmid was transfected into BMSCs, and osteogenic differentiation was induced. Proteins were extracted from the cells at different timepoints, and representative images were obtained after co-IP experiments with anti-UAP1 antibodies. G The V5 Ub overexpression plasmid was transfected into HEK293T cells, which were then cotransfected with Flag-TRIM25, Flag-TRIM25+Myc-UAP1 + HA-GLUL, or Myc-UAP1 + HA-GLUL overexpression plasmids. Subsequent Western blot images and quantitative analysis revealed the K48-linked ubiquitination of UAP1 in different cell groups. **P < 0.01 vs. other groups; *P < 0.05 vs. other groups.
