FIGURE 5.

Eltanexor treatment increases nuclear retention of IRF3 in HCMV infected or Poly (I:C) treated HFFs. (A) HFFs were infected with HCMV at a MOI = 1 and treated with Eltanexor (0.4 μM) or vehicle DMSO. At different time points, HFFs were fixed and immunostained with rabbit anti-IRF-3 antibody followed by FITC-conjugated goat anti-rabbit antibody (green). Nuclei were stained with DAPI (Blue). HCMV infected cells were indicated by mCherry (red). (B) Percentages of cells showing nuclear accumulation of IRF-3 in DMSO or Eltanexor treated HCMV-infected cells in (A) were calculated based on 5 random fields with more than 300 cells. The experiments were repeated 3 times. Data represent mean ± SEM, n = 3, **p < 0.01, ***p < 0.001. (C) Eltanexor treatment increases the nuclear accumulation of IRF-3 in poly (I:C) treated HFFs. Poly (I:C) was added the culture medium at a concentration of 20μg/mL. HFFs were fixed and immunostained for IRF-3 as described in (A). (D) Percentages of cells showing nuclear accumulation of IRF-3 in (C) were calculated based on 5 random fields. Data represent mean ± SEM; n = 5, ***p < 0.001. (E) Nuclear retention of viral tegument protein pp65 in Eltanexor treated HFFs at 72 hpi. HFFs were infected with HCMV at a MOI = 1, and treated with Eltanexor (0.4 μM) or vehicle DMSO. Cells were immunostained for pp65 (green). Nuclei were stained with DAPI (blue). HCMV infected cells were indicated by mCherry (red). The experiments were repeated 3 times.