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. 2003 Sep;122(3):333–348. doi: 10.1085/jgp.200308798

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Copyright © 2003, The Rockefeller University Press

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Figure 3. — Binding of [α³²P]8-N₃ATP to split CFTR channels. (A) Membranes from oocytes coexpressing Flag-3–835 CFTR and Flag-837–1480 CFTR (+), or noninjected controls (−), incubated with 5 or 50 μM [α³²P]8-N₃ATP at 0°C were irradiated with UV before washing, immunoprecipitation, and SDS-PAGE. Arrow indicates photolabeled band in the autoradiogram corresponding to CFTR half containing NBD1. (B) Immunoblots of proteins immunoprecipitated and run in same SDS-PAGE gel used for A, before transfer to nitrocellulose and blotting with anti-R domain antibody (left) that recognizes Flag-3–835-containing NBD1, or with anti–COOH-terminal antibody (right) that recognizes Flag-837–1480-containing NBD2.