Fig 1.

Surface distribution of ephrin receptors. Polarized MDCK cells grown on filter supports were fixed with 4% PFA and incubated with recombinant mouse EphB4/Fc, a soluble ephrin-B2/-B3 ligand fused to the Fc region of human IgG. Bound EphB4/Fc was stained with rhodamine- and AF 568-conjugated secondary antibodies (red). Adherens junctions were visualized after permeabilization by antibodies directed against E-cadherin and AF 488-conjugated secondary antibodies (green). (A and B) Confocal images representing apical, center, and basal horizontal sections through the cell monolayer (A) and a vertical section (B) are shown. (C) To control MDCK cell polarity, E-cadherin and the apical marker protein gp114 (canine CEACAM1) were detected after cell permeabilization by specific primary and according secondary antibodies. Bars, 10 μm.