Fig. 1

HIV-1 expressing a Gag-fusion protein triggers a type I IFN response in THP-1 cells. A: IFIT-1 reporter activity from monocytic THP-1-IFIT-1 cells transduced for 24 h with WT LAI (LAI), LAI packaged with 8.91 Gag (8.91 LAI), LAI expressing gag fused to luciferase and packaged with 8.91 Gag (Gag-LUC) or LAI expressing Gag fused to GFP and packaged with 8.91 Gag (Gag-GFP) (See Suppl. Figure 1) at 0.5, 1 or 2 U RT/ml. B-D: ISG qRT-PCR from PMA-treated THP-1 shSAMHD1 cells transduced for 24 h with LAI or Gag-LUC viruses at 0.125, 0.25 and 0.5 U RT/ml. E: CXCL-10 protein in supernatants from B-D (0.5 U RT/ml, ELISA). F: RT products from THP-1-IFIT-1 cells transduced for 24 h with 1 U RT/ml of the indicated viruses or transduced with boiled virus as a control. G: IRF reporter activity from monocytic THP-1 Dual cells transduced for 24 h with 1.5 U RT/ml LAI or Gag-LUC viruses, or stimulated with 1 ng/ml IFNβ as a control, in the presence of DMSO vehicle or 2 µM ruxolitinib. H, I: ISG qRT-PCR from monocytic THP-1 Dual cells transduced for 24 h with 1.5 U RT/ml LAI or Gag-LUC viruses, or stimulated with 1 ng/ml IFNβ as a control, in the presence of DMSO vehicle or 2 µM ruxolitinib. Data are mean ± SD from biological triplicates of a single experiment, representative of at least 3 repeats. Statistical analyses were performed using Student’s t-test, with Welch’s correction where appropriate, comparing each virus with WT LAI at the same dose (A-E) or pairs of samples -/+ ruxolitinib (G-I). *P < 0.05, **P < 0.01, ***P < 0.001