Figure 9

Effects of down-regulation or block of UTR with either an anti-UTR shRNA or the specific antagonist urantide on cell motility and invasion of RT112 cells. A: RT112 parental (CTR) or transiently transfected with a shRNA for UTR (shUTR), were plated in the top chamber of non-coated polyethylene teraphthalate (PET) membranes, treated or not with 100 nM urantide for 48 h and cell motility was evaluated as described in Methods Section. B: For in vitro invasion assays, RT112 cells were added to a Boyden chamber coated with Matrigel and cell invasion was evaluated as described in Methods Section. The migrating and the invading cells were stained with 0.25% crystal violet for 10 min and photographed under a microscope. C: The histogram shows the quantification of the migrating and invading cells measured with a spectrophotometer as OD, and the results are expressed as a percentage as compared to untreated RT112 parental cells. The experiments were performed three different times and the results are the mean of the obtained values. Bars, SDs. CTR, untreated RT112 cells; Sc, RT112 cells transfected with scrambled vector and cultured for 48 h; Sc + UR, RT112 cells transfected with scrambled vector and exposed for 48 h to 100 nM urantide; shUTR, RT112 cells transfected with shUTR and cultured for 48 h; shUTR + UR, RT112 cells transfected with shUTR and exposed to 100 nM urantide for 48 h. Asterisks indicate the statistical significance of the data (P <0.005).