Expression of EpCAM-YFP constructs in Esh2 lines depleted of EpCAM rescues Claudin-7 localization to the basolateral membrane.

Confocal images were acquired of cells 24 hours after high density plating on IBIDI chambers and 0 hours after removal of the chamber walls. The figure shows maximum intensity projections and orthogonal projections of confocal stacks of Esh2, EY, EIY and EEY cells stained for DNA, for YFP in the EpCAM-YFP fusion proteins and for Claudin-7. In the Esh2 line, Claudin-7 does not distribute along the basolateral membrane. In the EY, EIY and EEY expressing lines Claudin-7 localizes along the basolateral membrane. In the Esh2 line, greater contrast was applied post-processing to visualize the weaker Claudin-7 staining. In the EY, EIY and EEY the EpCAM-YFP fusion proteins localize along the basolateral membrane. Note, that Esh2 cells do not express any YFP-fusion protein and show no YFP-specific signal (see Fig 5C for anti-EpCAM staining of Esh2). Scale bar is 10μm.