Supplementary Figure 3: EPRS deficiency in mouse BMDMs reduces antiviral innate immune responses.
From: Infection-specific phosphorylation of glutamyl-prolyl tRNA synthetase induces antiviral immunity
(a) Immunoblot analysis of EPRS expression in BMDMs. BMDMs were transfected with non-targeting control siRNA (siCtrl) or siEPRS for 36 h. (b) Plaque assay to determine virus titers and (c) ELISA to measure IFN-β and IL-6 levels at 12 and 24 h post-infection. BMDMs were infected with PR8-GFP (MOI = 3) or VSV-GFP (MOI = 5) (b,c). (d) IFN-β and IL-6 levels measured in the culture supernatants from BMDMs treated with 40 μg of Poly(I:C). (e) Induction of Ifnb mRNA or IFN-related antiviral genes in virus-infected cells. RAW264.7 cells were transfected with siCtrl or siEPRS for 36 h, followed by infection with PR8-GFP (MOI = 1) for 12 h. The graphs show the -fold induction of the indicated genes after normalization against Gapdh. (f) Viral titer (determined by plaque assay) and (g) secreted IFN-β or IL-6 levels in cell culture supernatants after infection with HSV-GFP. BMDMs from Eprs+/+ and Eprs+/ – mice were infected with HSV-GFP (MOI = 2) (f,g). *P < 0.05, **P < 0.01, and ***P < 0.001 (Student’s t-test; b–d). Data are representative of three (b-d) or two (e-g) independent biological replicates with similar results (mean and s.d. of triplicate in b-g).
