Fig. 4. TRIM50 induced ubiquitous degradation of SNAIL by K-48 linked poly-ubiquitination.

a BEL7402 cells and HepG2 cells were transfected with TRIM50 plasmid and/or HA-UB plasmid as indicated, and further cultured for 24 h. The ubiquitous status of SNAIL was analyzed by co-IP assay. b HUH cells were co-transfected with Myc-tagged TRIM50 plasmid or TRIM50 truncation mutant (△RING) together with SNAIL expression plasmid, and further cultured for 24 h. The ubiquitous status of SNAIL was analyzed by co-IP assay. c BEL7402 cells and HepG2 cells were co-transfected with TRIM50 and HA-K-48-UB/HA-K-63-UB plasmids, and the ubiquitous status of SNAIL was analyzed by co-IP. d Cytoplasmic and nucleic fractions of HCC cells were prepared from HepG2 cells, and co-IP assay was used to detect the interaction between TRIM50 and SNAIL in different fractions of HCC cells. e The ubiquitous status of SNAIL protein in different fraction of HCC cells was also detected by co-IP. Lamin B1 was used as nuclear internal control, GAPDH was used as cytoplasm control, and β-actin was served as whole-cell loading control. Similar results were obtained in at least three independent experiments