MOV10 overexpression restricts the production of infectious retrovirus particles for a broad range of exogenous retroviruses. (A) MOV10 overexpression decreases HIV-1 virus production. HeLa or 293T cells were co- transfected with pHIV-1NL4-3 and increasing amounts of pT7-MOV10 as indicated or the pT7-Luc control. Virus concentration in the medium was determined by a p24Gag ELISA. Cell lysates were analysed by quantitative immunoblotting with anti-T7, anti- p24Gag and anti-Hsp90 antibodies. (B) Overexpression of MOV10 inhibits the infectivity of HIV-1 virions. The TZM-bl reporter cell line expressing a HIV-1 Tat inducible β-gal reporter gene was infected with equal amounts of virus normalised by the p24Gag concentration from each of the indicated samples. Cells were lysed and β-gal activity was measured to determine virus infectivity. (C) MOV10 overexpression inhibits the production of infectious SIVmac, MLV and M-PMV particles. For HIV-1 and SIVmac lentiviral vector production, 293T cells were co-transfected with p8.91, pCSGW and pVSV-G, or pSIV3+, pSIV-RMES4 and pVSV-G, respectively, together with pT7-MOV10 or pT7-Luc. 293T cells were infected with lentiviral particles and infectivity was determined by measuring the percentage of GFP-positive 293T cells by FACS. For MLV and M-PMV virion production, 293T cells were co-transfected with pNCS/FLAG, pMSCV/Tat and pVSV-G, or pMTΔE and pVSV-G, respectively, together with pT7-MOV10 or pT7-Luc. Infectivity was determined using TZM-bl cells. (D) Overexpression of MOV10 decreases the production of M-PMV virions. Cell lysates and sucrose cushion purified M-PMV virions were analysed by immunoblotting with anti p27Gag and anti-Hsp90 antibodies (* refers to non-specific bands). For (A), (B) and (C) results are normalised to the pLuc control, which is set at 100%. For (C) a single control bar set at 100% is graphed for simplicity. Values are the mean ± SD of 3 independent experiments.