Fig. 6

Mir-139-5p reversed EMT via down-regulating the expression of Notch1. (a and b) qRT-PCR analysis demonstrated that downregulation of Notch1 resulted in a reduction of Notch1, N-cadherin, Vimentin and Fibronectin mRNA expression and an elevation of E-cadherin mRNA expression. (c and d) Western blot analysis showed that the protein expression of Notch1, N-cadherin, Vimentin, Fibronectin, Snail-1 and Shh decreased, and the protein expression of E-cadherin increased in LN229 and U87 cells treated with Notch1 siRNA. (e and f) Notch1, E-cadherin, N-cadherin and Vimentin expression levels in the LN229 and U87 cells transfected with full-length Notch1 without the corresponding 3′-UTR or/and miR-139-5p were assessed by Western blotting. (g and h) Invasiveness of LN229 and U87 glioma cells transfected with full-length Notch1 without the corresponding 3′-UTR or/and miR-139-5p were estimated by transwell assays. This experiment was repeated three times. (× 50 magnification). (*P < 0.05. **P < 0.01. ***P < 0.001)