Figure 3: EpCAM opposes cadherin switching and stabilizes ENT1 at the cell surface.
A. Western blotting analysis of hENT1, EpCAM, N-cadherin, and E-cadherin expression following stable retroviral overexpression of EpCAM in PANC-1 and N+/E− cells. β-actin is used as an internal loading control. OE, overexpression. B. Immunostaining for EpCAM (green) and hENT1 (red) in PANC-1 control and PANC-1 overexpressing EpCAM. C. Z-intensity projection of compiled confocal micrographs show ENT1 (red) and EpCAM (green) localization throughout entire cells. Yellow transverse lines indicate position of slices where orthogonal view is shown at the bottom and to the right of the image and Z-projection 3D perspective view at 45° from horizontal; Bar, 10 μm. D. Immunostaining for EpCAM (green) and hENT1 (red) in N+/E− and N+/E− cells overexpressing EpCAM E./F. Transport of 3H-gemcitabine (1 μM) in PANC-1 and N+/E− cells stably expressing EpCAM conducted using sodium containing buffer. Bars, means ± SD (n=3). Statistical analyses were performed with two-tailed Student’s t-test. *P ≤ 0.05, **P ≤ 0.01