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. Author manuscript; available in PMC: 2018 Mar 1.
Published in final edited form as: J Immunol. 2017 Jan 25;198(5):1974–1984. doi: 10.4049/jimmunol.1600797

Figure 5. HCV ssRNA mediates monocyte differentiation.

Figure 5

Healthy monocytes were transfected with 5μg/ml of different regions of HCV ssRNA or control RNA and cultured for 7 days. (A) Cell surface expression of CD14, CD68, CD206 and CD163 were measured by flow cytometry. (B) TNFα, IL-1β, IL-10 and TGFβ levels were measured in culture supernatants by ELISA. (C) Viral concentrate consists of free HCV, ssHCV RNA and exosome-containing HCV. RNAse treatment can degrade the ss viral RNA, but not the enveloped HCV RNA in a dose dependent manner. Permeabilization and RNAse treatment of viral concentrate degrades both enveloped and non-enveloped viral RNA. (D) Viral concentrate was treated with 1U/ml of RNAse in the presence or absence of permeabilization buffer. The viral RNA levels were determined by quantitative RT-PCR method. The graph is represented as viral RNA copies/μl. (E) Monocytes were cultured in the presence of viral concentrate with or without treatment. Cell surface expression of CD14, CD68, CD206 and CD163 were measured by flow cytometry. (F) TNFα, IL-1β, IL-10 and TGFβ levels were measured in culture supernatants by ELISA. The data is represented as mean±SEM, n=3.