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. Author manuscript; available in PMC: 2010 Mar 16.
Published in final edited form as: Curr Biol. 2005 Apr 12;15(7):661–666. doi: 10.1016/j.cub.2005.02.051

Figure 3. Ty1 Gag Binds APOBEC3G Specifically.

Figure 3

(A) Western blot analysis of whole cell lysates from human 293T cells cotransfected with pK-GST (lanes 1 and 2), pK/HIVGAG-GST (lanes 3 and 4), or pK/TYA-GST (lanes 5 and 6) and with pK-h3G-HA (lanes 1, 3, and 5) or pK-βARR-HA (lanes 2, 4, and 6) by using anti-HA antibodies (top, input) or anti-GST antibodies (bottom). The fraction of each lysate that bound to glutathione-Sepharose beads was subjected to Western analysis with anti-HA antibodies (middle, bound).

(B) 293T cells were cotransfected with phA3G-HA together with pK/TY1GAG-GST (lanes 1 and 2) or pK/MS2-GST and pDM128/4XMS2 (lanes 3 and 4). Binding assays were performed as described in (A) except that the cell lysate was incubated at 37°C for 10 min in the presence (lanes 2 and 4) or absence (lanes 1 and 3) of 50 μg of RNase A, prior to addition of the glutathione-Sepharose beads.