Fig. 8
Gag processing requires both Pol and the viral genome. a Schematic structures of the four FFV genomes mATG, mATG-GC, mΔBBBB-GC and EO-GC. b 293T cells were co-transfected in 10 cm dishes with 2 µg of pBC-Gag-oPRE-based Gag constructs (wt, Q11A, H25A, R32A, G36A, R43A or L51A), 2 µg pMP-Pol-oPRE and 2 µg of the genome mATG. The viral genome was substituted by pcDNA as a negative control. Cell lysates were analyzed by SDS-PAGE. Positions of Gag p48 and p52, Pol precursor p127Pol and PR-RT-RN domain p65Pol are marked. c 293T cells were co-transfected in 10 cm dishes with 2 µg pBC-Gag-oPRE, 2 µg pMP-Pol-oPRE, 2 µg pBC-Env and 2 µg of each of four vector genomes. pcDNA was used as the negative control. Cell lysates and VLPs in supernatants were analyzed by SDS-PAGE. Positions of Gag p48 and p52, Pol precursor p127Pol and PR-RT-RN domain p65Pol are marked. d Viral RNAs in Gag particles were quantified by qRT-PCR. The figure shows an average of two independent experiments, with error bars indicating the range. Data is given as the relative number of cDNA copies compared to the RNA packaging-competent vector mATG, which was set to a value of 100 %