Fig. 3. Expression of trophectodermal and lineage specification markers in Ncad k.i. mutants. (A,A') Double immunolabeling of E3.5 embryos with antibodies against Oct4 (green) and Cdx2 (red) shows predominantly ICM localization of Oct4 and trophectodermal expression of Cdx2 in wt (A) and in mutant embryos (A'). Some outer cells are also Oct4-positive in mutant embryos (A'). (B,B') Monoclonal TROMA-1 antibody detects intermediate filament marker cytokeratin 8 in mutant (B') and wt (B) embryos. (C,C') Immunolocalization of ZO-1. In wt embryos ZO-1 localization is restricted to tight junctions (C). More extended and less punctate membrane localization of ZO-1 protein is seen in homozygous Ecad^{Ncad-GFP/Ncad-GFP} mutants (C'). (D,D') Polyclonal anti-ezrin antibodies were used to detect microvilli on the apical pole of the outer cells in compacted wt (D) and mutant (D') embryos. (E,E') Actin cytoskeleton was detected with Alexa-488-conjugated Phalloidin. Scale bar: 25 µm.