Fig. 5

NOSTRIN down-regulation potentiates the metastatic potential of HT29 cells. A Quantitative real time PCR analysis of Nostrin using RNA from HT29 cells transfected with either scramble or a cocktail of two Nostrin siRNAs (50 nM or 100 nM). B Quantitative real time PCR analysis of Vimentin (Vim) and Moesin (Msn) using RNA from HT29 cells transfected with either scramble or 50 nM Nostrin siRNA cocktail. GAPDH was used as an endogenous control for normalization. C Photo-micrographic images of invaded HT29 cells transfected with either scramble or 50 nM Nostrin siRNA cocktail from cell invasion assay. Scale bar: 100 μm, Magnification: 100X. D Invaded cell count per microscopic field. E Colorimetric quantification of the invaded cells (n = 3). F Representative photo-micrograph depicting wound closure ability of HT29 cells transfected with either scramble or 50 nM Nostrin siRNA cocktail. Magnification used, 50X. G Quantification of percentage wound closure 24 h post scratching from a minimum of 5 measurements in each experiment using three different biological replicates. H Western-blot analysis of EMT-associated proteins in control and NOSTRIN down-regulated HT29 cells. Full length blots are shown in Fig. S3. I Quantification of the proteins relative to GAPDH using three biological replicates using NIH ImageJ software. Error bars represent standard error of mean from three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001