Figure 3

Rab11 overexpression induced E-cadherin expression and cell migration in HT-29 cells. HT-29 cells were transfected with GFP-tagged Rab11 plasmid (a-c) or plasmid constructed with Rab11 shRNA (d-f). After 24 hr, cell lysates were collected and subjected to Western blot analysis for GFP-Rab11, E-cadherin and Rab11 expressions using anti-GFP, anti-E-cadherin and anti-Rab11 antibodies, respectively. GAPDH was used as the internal loading control. Quantifications of E-cadherin expression in GFP-Rab11 overexpressed cells (b) and in Rab11 knockdown cells (e) are shown from at least three independent experiments. Value = mean ± SD, * p < 0.05. Trans-well migration assay for Rab11 overexpression (c) and knockdown cells (f) were quantified. Value = mean ± SD were from three independent experiments, ** p < 0.01; *** p < 0.001. HT-29 cells grown on glass coverslips were transfected with GFP-Rab11; cells were then fixed and stained with anti-E-cadherin antibody, followed by staining with Cy3 conjugated secondary antibody. (g) GFP-Rab11-overexpressing cell migration out of the cell colony or (h) migration towards the center of the cell colony was observed by confocal microscopy. Arrowheads indicate cell membrane protrusion.