Fig. 5. Immunocytochemical analysis of occludin and ZO-1 in Caco-2 cells. (A,B) Brightfield and occludin labelling of subconfluent island after fixation showing typical TJ pattern of staining at cell borders. (C,D) Brightfield and ZO-1 staining of subconfluent island after fixation showing typical TJ pattern. (E-J) Brightfield and protein labelling of subconfluent islands immunolabelled prior to fixation. (E,F) Occludin labelling; note weak staining of the island periphery. (G,H) ZO-1 labelling showing absence of peripheral staining. (I,J) Negative control (secondary antibody only) showing absence of peripheral staining. (K,L) Confluent monolayer wounded to produce island and cultured for 30 minutes (K) or 1 hour (L) before labelling for occludin prior to fixation, showing weak staining around cells close to the wound edge (arrows). Bars, 20 µm.

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