Figure 2.

Paracrine communication from cells deficient in RIOK3 elicits diminished IFN response to infection with RVFV. (a) WT or RIOK3 KO cells were immune stimulated with poly(I:C) (1μg/mL), and media from these cells was transferred to identical plates of WT cells infected with RVFV (see diagram). IFNβ mRNA expression in the infected cells was measured by RT-qPCR. Cells treated with media from the WT cells elicited a stronger IFN response to infection than cells treated with media from RIOK3 KO cells. (b) Similarly, WT or RIOK3 KO cells were treated with TNFα at a concentration of 20 ng/mL, and media from these cells was transferred to identical plates of WT cells infected with RVFV (see diagram). Then, IL-8 and TNFα mRNA expression was measured by RT-qPCR. Cells treated with media from the WT or RIOK3 KO cells did not elicit statistically different inflammatory responses to infection, as measured by expression of IL-8 and TNFα. Fold induction in plots is relative to values of cells mock-infected and incubated with reduced serum media (negative controls). In (a,b), cells were grown in 6-well plates, seeded at a density ~0.3 × 10⁶ cells/well, and infected at a multiplicity of infection (MOI) ~2. Plots present the data as the mean value of 3 biological replicates +/− SEM. Student’s t-test: ** p < 0.01.