FIG 6.

Different protein tags or plasmid backbones affect the activity of NSP12. (A) HEK293T cells were transfected with pFlag-NSP12 and empty vector for 24 h. Flag-NSP12, and GAPDH were determined by Western blotting. (B) HEK293T cells were cotransfected with IFN-β luciferase reporter pIFN-β-Luc, pPRL-TK, and pFlag-NSP12 or empty vector for 24 h and then infected with SeV. After 10 h, relative luciferase activity was determined. (C) HEK293T cells were transfected with pFlag-NSP12 or empty vector for 24 h and then infected with SeV. After 10 h, IFN-β mRNA was determined by RT-qPCR. (D) HEK293T cells were transfected with NSP12 plasmid and empty vector for 12 h, 24 h, or 36 h. NSP12 mRNA was determined by RT-qPCR. (E) HEK293T cells were cotransfected with IFN-β luciferase reporter pIFN-β-Luc, pPRL-TK, and NSP12 plasmid or empty vector for 24 h and then infected with SeV. After 10 h, relative luciferase activity was determined. (F) HEK293T cells were transfected with NSP12 plasmid or empty vector for 24 h and then infected with SeV. After 10 h, IFN-β mRNA was determined by RT-qPCR.