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. 2018 Aug 3;14(8):e1007202. doi: 10.1371/journal.ppat.1007202

Fig 6. TLR3 mediates the MHV-68-induced SOCS1 production.

Fig 6

(A) BMMs were transfected with siRNAs against Tlr2, Tlr3, Tlr4, Tlr7, Tlr9 (si-TLR) or an irrelevant target (si-Control), respectively. Forty-eight hours post transfection, cells were infected with MHV-68 at MOI = 10. At 8 hpi, RNA and protein were then extracted. RT-qPCR was performed to determine SOCS1 mRNA levels and western blotting was performed to determine the protein expression levels of SOCS1 and the individual TLRs, respectively. The SOCS1 mRNA levels are expressed as values relative to the MHV-68 infected, si-Control transfected cells. (B) BMMs from Tlr3-/- and wildtype C57BL/6 mice were treated with poly(I:C) at 10 μg/mL for 1 and 8 hours, respectively. Total RNA was extracted and SOCS1 mRNA levels were determined by RT-qPCR. qPCR data are expressed as fold change in mRNA level compared to that in untreated WT cells. (C) BMMs from Tlr3-/- mice and wild type (WT) mice were infected MHV-68. At indicated hpi, cells were harvested to measure the SOCS1 mRNA levels by the RT-qPCR, and to measure the TLR3 and SOCS1 proteins levels by western blotting. qPCR data were expressed as fold change in mRNA level compared to that in uninfected WT cells. (D) BMMs were transfected with si-MyD88 or si-Control. Forty-eight hours post-transfection, the cells were infected with MHV-68. At 8 hpi, the SOCS1 mRNA levels and protein levels were determined. The SOCS1 mRNA levels are expressed as values relative to the MHV-68 infected, si-Control transfected cells. (E) BMMs from Myd88-/- mice and wildtype C57BL/6 mice were infected with MHV-68. At 8 hpi, total RNA was extracted and the mRNA expression of SOCS1 was determined by RT-qPCR. The SOCS1 mRNA levels are expressed as values relative to the MHV-68 infected WT cells. Results are representative of three independent experiments and are expressed as mean ± S.E.M., *p < 0.05.