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. 2019 Feb 11;15(2):e1007582. doi: 10.1371/journal.ppat.1007582

Fig 4. 14-3-3η is a critical chaperone protein for MDA5-mediated antiviral activity.

Fig 4

(A) IFNβ promoter activities of NTV and 14-3-3η K/D Huh7 cells during SeV infection or poly(I:C) stimulation. NTV and 14-3-3η K/D Huh7 cells were first transfected with pIFNβ-Luc, pCMV-rRL reporter plasmids and subsequently infected by SeV or simulated with poly (I:C) for 18 hours. The promoter activities of IFNβ were evaluated by dual luciferase assay. (B) NTV and 14-3-3η K/D Huh7 cells were mock-treated, infected with SeV, or transfected with poly(I:C) to monitor the expressions of IFNβ mRNA in these cells by quantitative RT-PCR. (C) NTV and 14-3-3η K/D cells were mock-transfected or transfected with 1 μg/mL poly(I:C) for 3 to 48 hours. The intracellular RNA from these cells was extracted and subsequently the IFNβ mRNA expression levels post poly(I:C) stimulation were determined by quantitative RT-PCR. (D and E) NTV or 14-3-3η K/D Huh7 cells were mock-treated or stimulated with poly(I:C) for 9 or 18 hours. The IFNβ and OAS1 mRNA expression levels were determined by quantitative RT-PCR. (F) NTV and 14-3-3η K/D Huh7 cells were mock-treated or infected with EMCV with different M.O.I. for 18 hours. The intracellular RNA from these cells was extracted and the IFNβ mRNA expression levels post EMCV infections were evaluated by quantitative RT-PCR. (*: p<0.05) (G) NTV and 14-3-3η K/D Huh7 cells were mock-treated or infected with EMCV for 18 hours. The intracellular EMCV viral RNA levels was determined by quantitative RT-PCR. (*: p<0.05).