Fig. 2: TRIM50 inhibited proliferation, colony formation, and invasion of HCC cells. | Cell Death & Disease

Fig. 2: TRIM50 inhibited proliferation, colony formation, and invasion of HCC cells.

From: TRIM50 suppressed hepatocarcinoma progression through directly targeting SNAIL for ubiquitous degradation

Fig. 2

a The basic protein levels of TRIM50 in BEL7402, SMMC7721, HepG2, and HUH7 cells were detected by western blot. b BEL7402 and HUH7 cells were transfected with TIRM50 expression plasmid or mock control, and western blot assay was performed to define the successful exogenous overexpression of TRIM50 in HCC cells. c BEL7402 and HUH7 cells were transfected with TRIM50 plasmid or mock control, and proliferation status of the transfected HCC cells was detected at 0 h, 12 h, 24 h, 36 h, and 48 h by CCK8 assay. d BEL7402, HUH7, and HepG2 cells were transfected with TRIM50 expression plasmid or mock control, and further cultured for 24 h before being transferred to six-well plates at the density of 1000 cells per well for colony formation assay. The clone formations were harvested after 14 days and the number of clone formation was counted. e, f After transfection with TRIM50 plasmid or mock control, transwell migration assay (e) and transwell invasion assay (f) were performed to investigate the migration and invasion capabilities of HCC cells. g HepG2 cells were transfected with siRNAs specifically targeting TRIM50 (Si-TRIM50-1&2, and Si-TRIM50-3), the cells transfected with random sequences (Si-NC) were used as mock control. The cells were further cultured for 24 h before being harvested and the block efficiency was measured by western blot. h HepG2 cells were transfected with siRNAs specifically targeting TRIM50 (Si-TRIM50-1&2) or its nonsense control (Si-NC), and proliferation status of transfected HCC cells were measured at 0 h,12 h, 24 h, 36 h, and 48 h by CCK8 assay. i HepG2 cells and SMMC7721 cells were transfected with Si-TRIM50-1&2 or Si-NC and further cultured for 24 h. The transfected cells were further transferred to six-well plates at the density of 1000 cells per well and allowed to grow for 14 days for colony formation assay. j, k After transfection of Si-TRIM50 or nonsense control (Si-NC) to HepG2 cells, transwell migratory assay (j) and invasion assay (k) of HCC cells were performed. Similar results were obtained in at least three independent experiments. *P< 0.05 and ***P < 0.001 for statistical analysis of the indicated groups