Figure 7

ASP-L -mediated inhibition of HIV-1 replication. Results of transient transfection experiments are shown in A to C. (A) HIV-1 RNA levels at the indicated time points with prior transfection of the ASP-L expression vector, pIRES-RSV-ASP-L, or a vacant vector into MAGIC-5A cells. The relative expression levels of HIV-1 gag RNA are presented relative to that of HIV-1 gag RNA expressed in mock-transfected cells set as 1.0. (B) Agarose gel electrophoresis of PCR products of semi-quantitative PCR of genomic DNA samples harvested at the indicated time points. ‘AL’ stands for pIRES-RSV-ASP-L transfected cells. (C) The levels of viral particle production in culture supernatants evaluated by RT assays. MAGIC-5A cells were transfected with the indicated amounts of the ASP-L expression vector before infection with HIV-1. (D–H) Prolonged inhibition of HIV-1 replication in ASP-L expressing Molt-4 clones. Three clones that stably express ASP-L and three clones transfected with the vacant vector were used. (D) The levels of virus production evaluated by RT assays. (E and F) HIV-1 RNA levels at 1 day (E) and 4 days (F) post infection (dpi) measured by qRT-PCR. (G) Agarose gel electrophoresis of PCR products of semi-quantitative PCR of genomic DNA samples at 1 and 4 dpi.. (H) The relative expression levels of ASP-L that were measured by qRT-PCR.