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Fig. 8. Threshold levels of E4-ORF1 expression are required to block TJ localization of PDZ proteins in MDCK cells. (A) High or low levels of E4-ORF1 protein expression by two MDCK lines. Extracts of sample buffer-lysed MDCK cells, MDCK-E4-ORF1-low cells and MDCK-E4-ORF1 cells (180 µg protein) were immunoblotted with E4-ORF1 or Erk1/2 antibodies. Erk1/2 served as a loading control. (B) The localization pattern of E4-ORF1 depends on its expression level in MDCK cells. MDCK-E4-ORF1-low cells or MDCK-E4-ORF1 cells were immunostained with E4-ORF1 antibodies. Scale bar: 40 µm. (C) E4-ORF1 must be expressed at a minimal threshold level to block TJ localization of PDZ proteins in MDCK cells. MDCK-E4-ORF1-low cells and MDCK-E4-ORF1 cells were immunostained with PATJ, ZO-2, SAP97 or ß-catenin antibodies. (D) E4-ORF1 protein levels progressively increase during an Ad9 infection of epithelial cells. A549 cells were infected with Ad9 virus for the indicated times, and extracts (80 µg protein) of sample buffer-lysed cells were immunoblotted with E4-ORF1 antibodies.





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