Fig. 3

Upregulation of Zeb1 mediates the RP11-induced dissemination of CRC cells. a. The expression levels of EMT-TFs in HCT-15 or HCT-8 RP11 stable overexpression and control cells were verified by western blot analysis. After transfection with si-NC or si-RP11 for 48 h, the expression levels of EMT-TFs in SW620 cells were verified by western blot analysis. b Zeb1 expression in subcellular fractions of HCT-15 RP11 stable overexpression and control cells was verified by western blot analysis. c IHC analysis of Zeb1-stained paraffin-embedded sections obtained from xenografts. d The mRNA expression levels of EMT-TFs in HCT-15 RP11 stable overexpression and control cells were verified by qRT-PCR. e After transfection with si-NC or si-Zeb1 for 48 h, the wound healing of HCT-15 RP11 stable overexpression and control cells was quantitatively analysed. f After transfection with si-NC or si-Zeb1 for 48 h, the EMT markers of HCT-15 RP11 stable overexpression and control cells were detected by western blot analysis. g After treatment with 100 μg/ml CHX for the indicated times, Zeb1 expression in HCT-15 RP11 stable overexpression and control cells was detected by western blot analysis (left) and quantitatively analysed (right). h & i Zeb1 in HCT-15 or HCT-8 RP11 stable overexpression and control cells was immunoprecipitated for the detection of ubiquitylation. Data are presented as the mean ± SD from three independent experiments. Bar = 200 μm. **p < 0.01 compared with control