Supplementary Figure 2: Knockdowns of components of the type III interferon pathway.

(A) Polarized T84 cells were treated with inhibitors of the JAK/STAT pathway: Fludarabine (STAT1), Pyridone 6 (JAKs) and AG490 (JAK2). Cells were subsequently infected with Reovirus, and expression of the ISG viperin was assessed by western immunoblotting. Huh7 cells (B) or JEG3 trophoblasts (C) were transfected with scrambled (Scr) or MAVS siRNA oligo. Efficient MAVS knockdown was monitored by western immunoblotting. (D) IRF3 depletion in Huh7 cells. ERK (E) or p38 (F) were knocked down in Huh7 cells. (G) IRF1 was knocked down in Huh7 cells. Cells were infected with SeV, inducing IRF1 in control cells transfected with a scrambled (Scr) siRNA oligo but not in cells transfected with an IRF1 siRNA oligo. (H) Huh7 cells were stably transduced with MAVS chimera localized on peroxisomes (Pex), mitochondria (Mito), both (WT) or neither (Cyto). MAVS transgenes include GFP whose expression is controlled by an IRES. Equivalent transgene expression was assessed by western immunoblotting against GFP. (I) Similar to H except JEG3 trophoblasts were transiently transfected with the indicated MAVS transgenes. Equivalent transgene expression was assessed by western blotting against GFP. (J) ERK1/2 was knocked down in MAVS-KO MEFs expressing MAVS-Pex.**, P < 0.01;****, P < 0.0001 (Student's t-test (D), One-way ANOVA (G)). Error bars represent mean ± SEM of triplicate readings for one experiment representative of 3.