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. 2021 Feb 26;13(3):367. doi: 10.3390/v13030367

Figure 2.

Figure 2

RIOK3 is involved in the signal transduction of the type I IFN response. (A) HEK293 cells and three RIOK3 KO cell lines were screened for RIOK3 protein expression by IP-Western blot. (B) HEK293 cells and RIOK3 KO cells were infected with MP-12 (MOI 1.0). The viral titer in the 24 hpi supernatant was quantified by plaque assay. (C,D) RT-qPCR targeting IFNB was performed on RNA from HEK293 cells and RIOK3 KO cells infected with MP12 (C) or treated with poly (I:C) (D) or 3p-hpRNA (E) for 18 hours. (F) RT-qPCR targeting IFNB was performed on RIOK3 KO cells transfected with a GFP (mock) or RIOK3 expression plasmid and treated with poly (I:C) 18 hours later. (G) Effect of RIOK3 KO on IFNB promoter activation. HEK293 cells and RIOK3 KO cells were co-transfected with pGL3-IFNB firefly reporter and phRL-CMV renilla control. Cells were stimulated with poly (I:C) or infected with MP-12 and the dual luciferase signals were measured after 18 or 48 h, respectively. In panels B–F, data are presented as mean +/− SEM from three biological replicates. Western blots in panels A is representative of duplicate experiments. Student’s t-test: ** p < 0.01, * p < 0.05.