Figure 6

MiR203 promotes MET. (a) Immunofluorescent images stained for E-cadherin (green), β-actin (red) and DAPI for nuclear localization of CDβ-Geo cells that have been treated with vehicle (control) or TGFb. (b) Total RNA was isolated from CDβ-Geo cells that have not (−) or have undergone EMT (+) and qRT-PCR analysis was performed to measure Zeb1 mRNA and mature miR203 expression levels. (c) Western blots of protein lysates from IMEC cells transiently transfected with miR-control or mature miR203 (60 nm) for 48 h were probed with anti-E-cadherin and anti-vimentin antibodies. (d) MDA-MB-231 cells were transiently transfected with miR-control or mature miR203 (60 nm) and plated for wound healing assay. At 90% confluence the cells were scratched and then observed for 48 h to measure closure of the wound. 10 × phase contrast microscopy of wound healing assay at 0, 24 and 48 h after scratch (c) and quantification of re-migration of the cleared space between experimental controls (e). All experiments were performed in triplicate (n=3) and data are means±S.D.