(a) SKIV2L and XRN1 were stably knocked down in primary macrophages, and the respective protein levels were evaluated by immunoblotting.
(b,c) Primary mouse macrophages, stably transduced as indicated with shRNA knockdown constructs, were treated with an HCV RIG-I ligand (b) or Poly (I:C) (c) and assessed for IFN-β production by quantitative RT-PCR.
(d–f) RIG-I ligand-stimulated macrophages with the indicated knockdown constructs were evaluated by quantitative RT-PCR for expression of IFNa4 (d), CXCL10 (e) and IL-6 (f).
(g,h) Macrophages were stimulated with DMXAA to activate STING directly, and with LPS to activate TLR4. *P < 0.05; **P < 0.001; ***P < 0.0005; ****P < 0.0001 (2-way ANOVA, b–f). Data are representative of at least three independent experiments with biological triplicates (mean + s.d., a–h).