Supplementary Figure 4: Dissociation of EPRS from the MSC upon viral infection.

(a,b) Viral infection induces dissociation of EPRS from the MSC component proteins. Lysates of RAW264.7 cells infected with PR8-GFP (MOI = 1) were subjected to immunoprecipitation with an anti-EPRS (a) or with an anti-KRS (b), followed by immunoblot analysis with anti-KRS, anti-MRS, anti-AIMP3, and anti-GAPDH (a), or with anti-EPRS and anti-AIMP3 (b), respectively. (c) Confocal microscopy of endogenous EPRS (red) and KRS (green) in HeLa cells infected with PR8 (MOI = 5) for 6 or 12 h. IFN-γ (1000 units/ml) treatment for 12 h was used for comparison. Cells were permeabilized with a lower dose of digitonin (20 μg/ml, 5 min) than used in Fig. 4b (25 μg/ml, 10 min). Scale bar, 10 μm (2 μm in magnified images). (d) Confocal microscopy of endogenous EPRS (red) and NSAP1 (green) in HeLa cells infected with PR8 (MOI = 5) for 6 or 12 h, or in cells treated with IFN-γ (1000 units/ml) for 12 h. Scale bar, 10 μm. Data are representative of three independent biological replicates with similar results (a-d).