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. 2019 May 15;10:1095. doi: 10.3389/fimmu.2019.01095

Figure 2.

Figure 2

Macrophage differentiation affected SFTSV replication. (A) THP-1 cells are differentiated into M1 macrophages by 16 h incubation with 150 nM PMA and followed by incubating with IFN-γ and LPS, or into M2 with interleukin 4 and interleukin 13 for 24 h. Cells were then fixed and immunolabeled for CD86 or CD206 using specific antibodies. Nuclei were stained with DAPI (blue). Bar = 50 μm. (B) Western blot analysis of SFTSV nuclear protein (NP) in M1-like or M2-like cells after infection with SFTSV for 24 h at an MOI = 1. (C) Immunofluorescence staining of viral dsRNA (green) in M1-like or M2-like cells after 24 h infection. Nuclei were stained with DAPI (blue). Bar = 50 μm. (D,E) Quantitative real-time-PCR analysis of replicative copy number of SFTSV in the extracellular (D) or intracellular (F) compartments of the infected M1-like or M2-like cells. (F) Schematic presentation of the transwell co-culture with SFTSV-infected Hela cells in the top well and polarizing THP-1 cells in the bottom well. (G) Immunofluorescence staining of viral dsRNA (green) in Hela cells. Nuclei were stained with DAPI (blue). Bar = 50 μm. All data are presented as the mean ± SEM of three independent experiments. (B,C) (**P < 0.01, ***P < 0.001).