FIGURE 5.
Release of HTLV-1 wt is inhibited in ITCH-silenced 293T cells. 3 × 105 293T derived cells stably silenced for the indicated UL were transfected with 0.2 μg of pCMVHT-1MΔenv or pCMVHT-1MΔenv K74R, 0.3 μg of pCRU5inGlucβ, 0.03 μg of pCMV-VSV-G. A, anti-ITCH immunoblot of cell extracts of the different lines showing the silencing of ITCH expression. Immunoblots of cell and pelleted VLP lysates show the reduced budding of wt HTLV-1 (B) and HTLV-1 K74R (C) in ITCH-silenced cells. D, relative infectivity of wt and K74R in stable cell lines silenced for the indicated UL. The infectivity in the cell line expressing a non-silencing shRNA was set at 100%. MA, monoclonal antibody.