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. Author manuscript; available in PMC: 2020 Jun 12.
Published in final edited form as: Cell Host Microbe. 2019 Jun 4;25(6):858–872.e13. doi: 10.1016/j.chom.2019.05.002

Figure 1. IFI16 impairs infectious HIV-1 production independently of type I IFN induction.

Figure 1.

(A-C) HEK293T cells were cotransfected with increasing doses of expression constructs for IFI16 (0.004, 0.02, 0.1, 0.5 and 2.5 μg) and constant quantities (2.5 μg) of the proviral HIV-1 NL4-3 construct. Cells and supernatants were harvested 40 hours post-transfection. Data show mean percentages (±SEM) relative to those detected in the absence of IFI16 (100%) and were derived from three experiments each performed in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001. (A) Infectious virus yield and p24 antigen levels in the HEK293T cell supernatants determined by TZM-b1 cell infection assay and p24 antigen ELISA, respectively. (B) Levels of different viral transcripts determined by qRT-PCR in multiplex reactions with GAPDH as control. (C) Expression of HIV-1 proteins, IFI16 and GAPDH in viral particles or cellular extracts was determined by Western blot.

(D) IFI16 does not induce IFN in HEK293T cells lacking STING expression. HEK293T cells were transfected with the indicated constructs (1 μg) or infected with Sendai virus (SeV). 40 hours later, cells and supernatants were harvested. Cells were used for Western blot analysis and supernatants were used to stimulate HL116 IFN-reporter cells in triplicates. An IFNα standard curve was used for comparison. After 8 hours, luciferase expression in the HL116 cells was determined. Shown are mean values from triplicates ± SD. See also Figure S1.