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Fig. 3. E4-ORF1 binds endogenous PATJ and redistributes it into detergent-insoluble complexes in MDCK cells. (A) Endogenous PATJ co-immunoprecipitates (IP) with E4-ORF1 in MDCK cells. Extracts (900 µg protein) of RIPA buffer-lysed MDCK cells, MDCK-E4-ORF1 cells and MDCK-IIIA cells were immunoprecipitated with E4-ORF1 antibodies, and recovered proteins were immunoblotted with PATJ and E4-ORF1 antibodies. (B) E4-ORF1 causes PATJ to redistribute into detergent-insoluble complexes in MDCK cells. Detergent-soluble or detergent-insoluble fractions of MDCK cells, MDCK-E4-ORF1 cells and MDCK-IIIA cells were prepared as described in the Materials and Methods, and an equivalent amount of each fraction was immunoblotted with PATJ or E4-ORF1 antibodies. An immunoblot assay of total cell extracts with PATJ antibody shows equivalent amounts of PATJ in the three lines (bottom panel). Vertical lines indicate the boundary between spliced lanes from the same gel.





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