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MDCK cells were internalized with pIgA-R ligand-Alexa488 and pIgA-R ligand-Cy3, for 4h at 17degrees C, from the apical and basolateral PM, respectively. Fluorescence confocal images were taken at ~3.5microns below the apical PM to evaluate FRET.

Schematic representation of a polarized MDCK epithelial cell, showing apical and basolateral PM. The enclosed star is the basolaterally internalized Cy3-pIgA-R-ligand complex. The open star is the apically internalized Alexa488-pIgA-R-ligand complex. Co-localization occurs in the apical endosome in the sub-apical region.