Figure 5.

N-cadherin was translocated from the plasma membrane to the intracellular compartment in response to CS-E. A, HeLa cells transiently expressing N-Cad(FL)-EGFP were incubated in the presence (panels c and d) or absence (panels a and b) of CS-E and were then fixed and permeabilized for immunofluorescence analysis of EGFP (green). Nuclei were counterstained with Hoechst (blue). B, HeLa cells transiently expressing N-Cad(FL)-EGFP were treated overnight with CS-A (panels b and b′), CS-C (panels c and c′), or CS-E (panels d and d′). The areas within the dotted white squares (panels b–d) are magnified and shown in panels b′–d′. Arrowheads indicate N-Cad(FL)-EGFP translocated from the cell surface into the intracellular compartment. C, HeLa cells transfected with the expression vector of N-Cad(FL)-EGFP were incubated in the presence (+CS-E) or absence (−CS-E) of 25 μg/ml CS-E overnight. Cells were stained with anti-LAMP-1 antibody (panels b and m), anti-EEA-1 antibody (panels e and q), anti-GM130 antibody (panels h and t), or anti-caveolin-1 antibody (panels k and w) and Alexa594-labeled second antibody (red). GFP florescence is shown in the left columns, and merged images in which nuclei were counterstained with Hoechst (blue) are shown at right. Magnified images of the areas within the white boxes in panels o, r, u, and x are shown in panels o′, r′, u′, and x′, respectively.