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. 2019 Aug 26;116(37):18619–18628. doi: 10.1073/pnas.1909314116

Fig. 6.

Fig. 6.

IRF1 regulates cytokine production during HCMV replication. (A–C) HFF cells were treated with siCon, siRoq, or siRoq+siIRF1 and infected with HCMV (MOI = 2), followed by harvesting at 72 hpi. (A) Protein levels of Roquin, IRF1, and viral genes were measured by immunoblot. (B) Cell-free virus in the supernatant was titrated by limiting dilution analysis. Data represent mean ± SEM, n = 3; ***P < 0 .001 (siCon versus siRoq; siRoq versus siRoq+siIRF1) according to two-way ANOVA with Dunnett’s multiple comparisons test. (C) Cytokine mRNA levels were measured by qRT-PCR (mean ± SEM, n = 3; *P < 0.05; **P < 0 .01; ***P < 0 .001 according to two-tailed Student’s t test; NS, not significant). (D) HFF cells were transfected with ASOs (at the indicated concentrations) targeting the Roquin-binding site on IRF1 mRNA, and IRF1 levels were measured by immunoblot. (E) HFF cells were transfected with ASOs and infected with HCMV (MOI = 0.1). At 24 hpi, protein levels of IRF1 and viral gene (UL44) were measured by immunoblot.