Previously published transcript sequences in the NCBI Entrez Nucleotide database: Ifit2, NM_008332; Ifit1, NM_008331; Ifit3, NM_010501; Ifnb1, NM_010510; Ifnar1, NM_010508. Figure S1 Survival of wt and Ifit2 2/2 mice after infection with low EMCV dose (25 pfu). Statistical significance of survival differences is indicated by p-value; n.s., not significant. (PDF) Figure S2 Enhanced ISG and IFN-b induction in intranasally VSV-infected Ifit2 2/2 brain regions. IFN-b-, and Ifit3/2/1 mRNA levels in different regions of brains of uninfected or VSV-infected wt and Ifit2 2/2 mice at 6 d.p.i., plotted as mean+SD. n = 4 mice per infected group; ND, not done. Infections were intranasal with 4610 2 pfu of VSV. (PDF) Figure S3 Gene induction in brains after VSV or EMCV infections. A, mRNA levels of select genes in brains (without OBs) of uninfected or intranasally VSV-infected wt and Ifit2 2/2 mice at 6 d.p.i., plotted as mean+SD; n = 3 mice per infected group; infection was intranasal with 4610 2 pfu of VSV. B, mRNA levels of select genes in brains (without OBs) of uninfected or intracranially VSV-infected wt and Ifit2 2/2 mice at 24 h post injection, plotted as mean+SD; n = 4 mice per infected group; infection was intracranial injection with 10 pfu of VSV. C, Ifit2, Ifit1, IFN-b and EMCV RNA levels in brains 4 days after EMCV infection (5610 2 pfu, n = 3 mice per infected group). (PDF) Figure S4 Region-selective induction of apoptosis in brains of intranasally VSV-infected Ifit2 2/2 mice. Ifit2 2/2 mice were i.n. infected with 4610 2 pfu of VSV; at 6 d.p.i., adjacent sections of fixed brains were labeled to detect apoptotic cells (TUNEL) or VSV P protein (immunohistofluorescence), n = 2 mice; only few regions such as striatum show positive TUNEL; infected wt brains and uninfected control brains of either genotype did not show appreciable signals, hence data not shown). (PDF) Figure S5 VSV yields from infected wt and Ifit2 2/2 MEF. Immortalized MEF were treated for 16 h with 10 U/ml IFN-b and infected with VSV at moi 10. 12 hours after infection, virus yields were determined by plaque assay. Results are plotted as mean+SD on log scale, representing one of two independent experiments. (PDF) Figure S6 Murine Ifit2 protein does not bind ppp-RNA. Single-stranded radiolabeled VSV leader RNAs (nt 1-18) with either 59-triphosphorylated or free 59-hydroxyl-ends (ppp-RNA or HO-RNA) were in vitro incubated with purified murine Ifit1 ( = P56) or Ifit2 ( = P54) proteins; formation of protein/RNA complex was detected by electrophoretic mobility shift assay. (PDF)