Fig. 3.

IFIH1 or MAVS knockdown prevents ISG15-induction by HIV-1 in macrophages. (A) CD14⁺ monocytes were transduced with shRNA-puromycin resistant vectors targeting IFIH1, MAVS, or LUC control, as indicated. Cells were selected with puromycin for 3 d, and differentiated into macrophages in the presence of GM-CSF. Cells were transduced with HIV-1-GFP for 3 d and then assessed by flow cytometry for intracellular ISG15. Plots show relative ISG15 signal in the indicated knockdown cells, with and without HIV-1-GFP transduction (mean ± SEM, n = 4 donors). P values were determined by one-way ANOVA with Tukey’s multiple comparisons test. (B) Quantification of GFP⁺ cell populations in donors used in the experiments for A (mean ± SEM, n = 4 donors). P values were determined by the two-tailed, paired t test. (C) As in A, except that macrophages were infected with either encephalomyocarditis virus (EMCV) or Sendai virus (SeV), instead of with HIV-1-GFP (mean ± SEM, n = 4 donors). P values were determined by one-way ANOVA with Tukey’s multiple comparisons test.