FIG. 2.
TRIM5αrh and TRIM5αhu mutants and retroviral restriction. (A) The primary sequences of the B-box 2 domains from TRIM5αrh and TRIM5αhu are aligned. Residues depicted in red are predicted to be exposed on the surface of the B-box 2 domain. The asterisks indicate residues that were changed by site-directed mutagenesis. wt, wild type. (B) Cf2Th cells were transduced with the LPCX vector expressing HA-tagged wild-type and mutant TRIM5αrh proteins. Stable cell lines were selected with 5 μg/ml of puromycin. Cell lysates were analyzed by Western blotting using antibodies against HA (Anti-HA) and β-actin. (C and D) Cf2Th cells expressing the indicated wild-type and mutant TRIM5αrh proteins were incubated with HIV-1-GFP, and GFP-positive cells were measured by fluorescence-activated cell sorting. The results of two independent experiments were similar, and the results of one experiment are shown.