Figure 4.

miR-106b directly target Prrx1 and formed a negative feedback loop with TGF-ß1 co-regulated Prrx1. A. The expression levels of Prrx1 in SW480 cells after inhibition of miR-106b or in SW620 cells overexpression of the same miRNAs were detected by western blot. B. Prrx1 3’UTRs are targets of miR-106b. PLuc-Prrx1 containing a wild-type or mutated Prrx1 3’UTRs (indicated as WT or MUT on the X-axis), were transfected into SW620 or SW480 cells. Relative repression of firefly Luciferase expression was standardized to a transfection control. The reporter assays were performed three times with essentially identical results. ***P<0.001. C. The cells migration capacity was measured by transwell in SW480 cells co-transfected with anti-miR-106b and siRNA/Prrx1. D. E-cadherin was examined by immunofluorescence in SW480 cells co-transfected with anti-miR-106b and siRNA/Prrx1. E. TGFB1 was examined by western blot after miR-106b over expression or down-regulation. F. The expression of Prrx1 was analyzed by a qRT-PCR array after the cells treatment with TGF-ß1. *P<0.05. G. The expression of miR-106b was analyzed by a qRT-PCR array after the cells treatment with TGF-ß1. **P<0.01. H. The expression of epithelial marker E-cadherin after co-treated with TGF-ß1 and siPrrx1 in SW620-lenti-miR-106b cells was examined by immunofluorescence. Cells were stained with E-cadherin (E-Cad) antibodies, and counter stained with DAPI.