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Fig. 7. Overexpression of myc-Pals1 or VSV-G Crb3 does not redistribute PATJ. (A) Caco2 cells were transfected by electroporation with a plasmid containing the cDNA of Pals1 tagged with a myc epitope and grown on filters for 3 days. Images are confocal microscopic Z-sections of cells processed for immunofluorescence as described in Materials and Methods and labeled with affinity-purified antibodies against PATJ (in blue) and myc (9E10) (in red). Myc-Pals1 is accumulated on the apical membrane (arrow) whereas PATJ is mainly at the level of tight junctions (arrowheads). (B,C) Overexpression of VSV-G-Crb3 leads to mislocalization of aPKC (B) but not of PATJ (C). Caco2 cells were transfected by electroporation with a plasmid containing the cDNA of hCrb3 tagged with a VSV-G epitope on the extracellular domain and grown on filters for 3 days. Images are confocal microscopic Z-sections of cells processed for immunofluorescence and labeled with affinity-purified antibodies against PATJ or aPKC (in blue) and VSV-G (P5D4) (in red). VSV-G-Crb3 is distributed in an unpolarized fashion when overexpressed and leads to mislocalization of aPKC (arrows) but not of PATJ (arrowheads). Bars, 10 µm.
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