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Fig. 6. Crb3 is present in an EEA-1 positive compartment in PATJ KD cells. Confocal microscopic Z-sections (A) and microscopic X-Y sections (B) of control (CT Cl 8) and PATJ KD clones (Cl 12 or 4) of Caco2 cells grown on filters and labeled with antibodies against EEA-1 (in red) and affinity-purified antibodies Crb3 (in blue). EEA-1 is found in early endosomes mostly accumulated below the apical membrane (arrowheads) and overlaps strongly with Crb3 in Cl 4 or 12 (arrows). In control cells, Crb3 is also found in small vesicular structures in the cytoplasm. (C) Immunogold labeling of PATJ KD Cl 4 cells prepared by cryosubstitution. Ultra-thin sections were labeled with antibodies against EEA-1 (15 nm gold particles) and affinity-purified Crb3 antibodies (6 nm gold particles). Crb3 labeling (arrowheads) is found at the plasma membrane and in sub-apical vesicles, some of which are also labeled with EEA-1 (arrows). ap, apical membrane; mv, microvilli. Bars, 10 µm (A,B); 100 nm (C).
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