Fig. 2.

Analysis of virus infectivity. Luciferase gene-transducing HIV viruses were produced by cotransfection of HEK 293T cells with a VSV-G expression vector plus WT and H62 mutant HIVLuc constructs (A) or the indicated HIV variants that derived from selection of H62A or H62F second-site revertants (B). At 3 days posttransfection, virus-containing medium supernatants were collected, filtered through 0.45-μm sterile filters, and used to infect HiJ cells. At 3 days postinfection, HiJ cells were collected and processed for luciferase assays. Results are depicted as normalized luciferase activity signals in infected cells, relative to signals from cells infected with WT HIVLuc virus in parallel: they are not corrected for variations observed in virus particle release. Results are derived from 46 (WT), 16 (H62A and H62F), 2 (H62C and H62F/G208A), 3 (H62K), 4 (H62W and H62Y), 5 (G208A), 8 (G208R and H62F/SP1P10L), and nine (H62A/G208R) separate experiments. Standard deviations are as shown.