HIV-1 association with rhTRIM5α cytoplasmic bodies requires the rhTRIM5α B30.2 domain essential for recognizing HIV-1 capsids and restriction. (A) CRFK cells stably expressing HA-tagged wild-type (wt) rhTRIM5α or rhTRIM5αΔpatch were immunostained with anti-HA antibody, and protein localization was visualized microscopically. The provided image is one z section from a deconvolved z-stack image. (B and C) Each cell line was also infected with VSV-G–pseudotyped HIV-1 virions dually labeled with S15-mCherry and GFP-Vpr in a synchronized infection. Cells were fixed either 90 min after infection (B) or 4 h after infection in the presence of MG132 (C), immunostained with anti-HA antibody, and imaged. The association of both unfused (S15⁺) and fused (S15⁻) HIV-1 virions with the HA-positive cytoplasmic bodies was examined in both cell lines. The mean results from three independent experiments are graphed, counting a total of 1,716 (B) and 3,381 virions (C). Error bars represent the SEM from three experiments.