HIV-1 VLP encapsidation and late restriction activities of human TRIM proteins.

<p>(A) Immunoblot analyses of C-terminally HA-tagged human TRIM proteins (1, 5α, 6, 18, 21, 22 and 34) with or without codon-optimized HIV-1 Gag-Pol expression plasmid (pH-G/P) are shown. TRIM5αrh was used as a positive control for incorporation. Top three panels show proteins harvested from producer cell lysates, immunoblotted against TRIM (anti-HA), HIV-1 (anti-p24; 183-5C-H12, 1∶1000 and AG3.0, 1∶500) or actin (anti-β-actin), while the bottom two panels show immunoblots of VLPs harvested from respective producer cells against TRIM (anti-HA) or HIV-1 (anti-p24; 183-5C-H12, 1∶1000 and AG3.0, 1∶500). FL designates approximate full-length TRIM size, and p24 designates HIV-1 p24 capsid size. (B) 293T cells were co-transfected with 1.0 µg of TRIM-expressing plasmid: TRIM5αrh, TRIM1, TRIM5αhu, TRIM6, TRIM18, TRIM21, TRIM22 and TRIM34, with 0.1 µg of lentivirus proviral plasmid: pNL4-3, p89.6, pROD10 or pSIV_MAC1A11. Viral titers were determined in GHOST(3)R3X4R5 indicator cells and reported as infectious units per ml (IU/ml). Error bars indicate one standard deviation.