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. 2001 Feb;12(2):463–473. doi: 10.1091/mbc.12.2.463

Figure 4.

Figure 4

In vitro binding assays. (A) μ2 binds to the N-terminal tyrosine-based motif in Nef from SIVmac239. Shown is an autoradiogram of the binding of hybrid GST-Nef proteins from SIVmac239 (GST-SIV Nef and GST-SIV Y28A) and HIV-1 (GST-SF2 and GST-NL4–3 Nef) to the in vitro translated μ2 chain from AP-2 (top). Input of the GST proteins is shown in the lower panel. (B) V1H binds to the flexible loop of Nef from SIVmac239. Shown is a liquid yeast two-hybrid assay testing the interaction of wild-type and mutant Nef proteins from SIVmac239 with V1H. The β-galactosidase activity is expressed in relative light units (RLU). (C) Similar internalization rate of V1H and Nef. Kinetic internalization assay of CD8, CD8-SIV Nef, and CD8-V1H chimeras in 293-T cells.