Supplementary Figure 2: Antiviral effects of EPRS in EPRS-deficient or EPRS-overexpressing immune cells.
From: Infection-specific phosphorylation of glutamyl-prolyl tRNA synthetase induces antiviral immunity
(a) Immunoblot analysis of EPRS expression. (b) Viral titer after infection with HSV-GFP (MOI = 1). RAW264.7 cells were transfected with non-targeting control siRNA (siCtrl) or siEPRS (a,b). (c) Fluorescence microscopy images, (d) virus replication, and (e) secreted IFN-β or IL-6 levels in 293T cells transfected with siCtrl or siEPRS for 36 h, followed by infection with VSV-GFP (MOI = 0.0001). (f) Immunoblot analysis of EPRS expression. (g) Fluorescence microscopy images, (h) PR8 titer, and (i) secreted IFN-β or IL-6 level in cells infected with PR8-GFP (MOI = 1). (j) Fluorescence microscopy images, (k) VSV titer, and (l) secreted IFN-β or IL-6 level in stable EPRS-deficient cells infected with VSV-GFP (MOI = 0.5). RAW264.7 cells were transduced with non-targeting control shRNA (shCtrl) or EPRS shRNA (shEPRS), followed by selection with puromycin (f–l). (m) Immunoblot analysis of EPRS expression. (n) Fluorescence microscopy images, (o) VSV titers, and (p) secreted IFN-β or IL-6 level in EPRS-overexpressing cells infected with VSV-GFP (MOI = 0.5). RAW264.7 cells were transfected with a FLAG-tagged empty vector (Ctrl) or with EPRS-FLAG (EPRS) plasmids, followed by selection with puromycin (m–p). Scale bars, 100 μm (c,g,j,n). *P < 0.05, **P < 0.01, and ***P < 0.001 (Student’s t-test; d,e,h,i,k,l,o,p). Data are representative of three (a-p) independent biological replicates with similar results (mean and s.d. of triplicate in b,d,e,h,i,k,l,o,p).
