FIG 7.

The infectivity of impairment-resistant HIV-1 AD8 Env does not circumvent the SERINC5-induced production of proinflammatory cytokines in MDMs. (A, upper panel) Relative infectivity of HIV-1 AD8 ΔNef particles produced in 293T cells transfected with a SERINC5 expression plasmid or a control vector. Particle infectivity was measured by infecting TZM-bl reporter cells and normalized to the amount of RT activity of the respective virus stock as determined by SG-PERT assay. (Lower panel) Representative Western blot analysis of these virions for presence of SERINC5.HA and HIV-1 p24. (B) Schematic of experimental flow (as in Fig. 2B, but with infection with HIV-1 AD8). (C) Relative intracellular levels of p24 as quantified by immunofluorescence (with ΔNef set as 100% and the percentage of p24⁺ cells ranging from 0.6% to 7% between donors). Shown are data points from cells of individual donors, with the mean of all donors indicated by a black line. (D and E) Quantification of IL-6 (D) and IL-8 (E) concentrations in the supernatant of the MDMs analyzed in panel C. NI, noninfected. The fold change between the ΔNef ± S5 conditions is indicated. Statistics (Student's t test): n.s., nonsignificant; **, P < 0.01.