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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
HEP_24189_sm_suppinfofigure1.tif463KSupporting Figure 1. Gene induction profiles following a single injection of IFN-alpha;, -β, -γ, and -λ in mouse liver and gut. C57Bl/6 mice were injected subcutaneously with increasing doses of murine IFN-α, -β, -γ and -λ and sacrificed 1 hour after injection. IFN doses were: 300, 1500 and 7500 pg/g bodyweight IFN-α; 100, 250 and 500 IU/g bodyweight IFN-β; 50, 100 and 500 ng/g bodyweight IFN-λ and 20, 100 and 500 IU/g bodyweight IFN-γ. Graphs show quantitative RT-PCR analysis of STAT1 (A, B), USP18 (C, D) and PKR (E, F) mRNA expression in liver (A, C, E) and small intestine (B, D, F). The data are plotted as the amount of STAT1, USP18 or PKR mRNA relative to RPL19 mRNA (mean with SEM).
HEP_24189_sm_suppinfofigure2.tif8537KSupporting Figure 2. IFN-β-induced signaling in the mouse liver is not abrogated after pre-treatment. (A, B) C57Bl/6 mice were injected subcutaneously with different murine IFNs and sacrificed 1 hour or 16 hours later, or injected again 16 hours after the first administration and then sacrificed 1 hour after the second injection. IFN doses were 300 pg/g bodyweight IFN-α, 500 IU/g bodyweight IFN-β, 50 ng/g bodyweight IFN-λ or 100 IU/g bodyweight IFN-γ. (A) Quantitative RT-PCR analysis of SOCS1 mRNA expression in the liver samples. The data are plotted as the amount of SOCS1 mRNA relative to RPL19 mRNA (mean with SEM). (B) Liver samples were subjected to immunoblotting for tyrosine phosphorylation of STAT1 and β-actin. Density of phospho-STAT1 and β-actin bands was measured using ImageJ software. The graph plots mean (with SEM) of the ratio of phospho-STAT1 and β-actin band densities.
HEP_24189_sm_suppinfofigure3.tif8537KSupporting Figure 3. IFN-β-induced signaling in the mouse kidney and lung is not abrogated after pretreatment. C57Bl/6 mice were injected subcutaneously with different murine IFNs and sacrificed 1 hour or 16 hours later, or injected again 16 hours after the first administration and then sacrificed 1 hour after the second injection. IFN doses were 300 pg/g bodyweight IFN-α, 500 IU/g bodyweight IFN-β, 50 ng/g bodyweight IFN-λ or 100 IU/g bodyweight IFN-γ. Kidney (A) and lung (B) samples were subjected to immunoblotting for tyrosine phosphorylation of STAT1 as well as STAT1 total protein and β-actin as a loading control.
HEP_24189_sm_suppinfofigure4.tif251KSupporting Figure 4. Repeated administration of IFN-λ does not lead to the reduction of the signaling in the mouse gut. C57Bl/6 mice were injected subcutaneously with different murine IFNs and sacrificed 1 hour or 16 hours later, or injected again 16 hours after the first administration and then sacrificed 1 hour after the second injection. IFN doses used were 300 pg/g bodyweight IFN-α, 500 IU/g bodyweight IFN-β, 50 ng/g bodyweight IFN-λ or 100 IU/g bodyweight IFN-γ. Small intestine samples were subjected to immunoblotting for tyrosine phosphorylation of STAT1 and β-actin. Density of phospho-STAT1 and β-actin bands was measured using ImageJ software. The graph plots mean (with SEM) of the ratio of p-STAT1 and β-actin band densities.

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