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Fig. 7. PI 3-kinase activation by E4-ORF1 is not required to block TJ localization of PDZ proteins in MDCK cells. (A) Endogenous ZO-1 fails to associate with TJs in MDCK cells expressing mutant IIB E4-ORF1. Cells of the MDCK-IIB pool were immunostained with ZO-1 (middle panel) or E4-ORF1 (left panel) antibodies. The combined images are shown in the right panel (merge). Scale bars: 40 µm. (B) Endogenous PATJ and ZO-2 also fail to associate with TJs in MDCK cells expressing mutant IIB E4-ORF1. Cells of the MDCK-IIB pool were immunostained with ZO-1, PATJ, and ZO-2 antibodies. The top three panels represent the same field of cells showing immunostaining of ZO-1, PATJ, or the merge of the latter two images (merge). The bottom three panels represent the same field of cells showing immunostaining of ZO-1, ZO-2, and the merged images (merge). Dashed lines outline groups of cells inferred to express E4-ORF1 based on the results shown in (A).