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. 1997 Mar 3;185(5):909–920. doi: 10.1084/jem.185.5.909

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Pulse-chase analysis and immunoprecipitation of nef. P815 cells infected with vaccinia vectors expressing β-gal (lane 1), nef (lanes 2–5), UbMNef (lanes 6–9), and UbRNef (lanes 10–13) were pulse labeled with ³⁵S-Met and ³⁵S-Cys for 30 min and then chased with an excess of unlabeled Met and Cys for 0 min (lanes 1, 2, 6, 10), 15 min (lanes 3, 7, 11), 60 min (lanes 4, 8, 12) or 240 min (lanes 5, 9, 13), followed by extraction, immunoprecipitation, and electrophoretic analysis of nef and MHC class I expression (see Materials and Methods).