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Fig. 1. E4-ORF1 directly binds to PDZ proteins expressed in MDCK cells. (A) E4-ORF1 directly binds to MDCK cell-derived PDZ proteins having gel mobilities similar to the known E4-ORF1-interacting PDZ proteins MAGI-1, ZO-2 and SAP97, but not MUPP1, expressed in TE85 cells. The detection of an unknown 220 kDa protein (p220) is indicated. Extracts (1.2 mg protein) of RIPA buffer-lysed TE85 and MDCK cells were subjected to pulldown reactions with the indicated GST fusion proteins, and recovered proteins, immobilized on a membrane, were blotted with a radiolabeled GST-E4-ORF1 protein probe. Proteins were detected by autoradiography. A vertical line indicates boundary between spliced lanes from the same gel. (B) MDCK cells lack detectable MUPP1 expression. Extracts (100 µg protein) of RIPA buffer-lysed TE85, CREF and MDCK cells were immunoblotted with MUPP1 antiserum. TE85 and CREF cells represented positive controls for MUPP1 expression in this experiment.





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