FIG. 8.

Higher-order self-association of TRIM5α_rh B-box 2 mutants. 293T cells were transfected with plasmids expressing the indicated wild-type or mutant TRIM5α_rh proteins with a FLAG or an HA epitope tag. Cells expressing wild-type and mutant TRIM5α_rh proteins were lysed 48 h after transfection. (A to D) The cell lysates were mixed, and the indicated mixtures were used for immunoprecipitation by an antibody directed against the FLAG epitope, as described in Materials and Methods. Elution of the immunocomplexes was performed with a FLAG tripeptide and analyzed by Western blotting with anti-HA and anti-FLAG antibodies. WB, Western blot; IP, immunoprecipitation. (E) The higher-order self-association and HIV-1-restricting ability, determined as described in the Table 2 footnotes and in Materials and Methods, of each TRIM5α_rh variant were compared. The means of the values for higher-order self-association within each restriction group are indicated by horizontal bars. The Spearman rank correlation coefficient, r_s, is 0.862, with a 95% confidence interval of 0.665 to 1.0 (two-sided P value of <0.001).