Fig. 7.

IL-10R signaling does not directly inhibit IFN-γ-induced STAT1 function. (A) A heatmap shows the Z-normalized expression of each of 30 leading edge genes from a previously defined gene set of IFN-γ-induced genes in bone marrow-derived macrophages⁵⁵ that was found by GSEA analysis to be significantly enriched in comparisons between WT and Il10ra^−/−, WT and WT_mix, and WT and Il10ra^−/−_mix colonic macrophages. Gene set enrichment false discovery rate Q values are shown for the comparison of all experimental groups; (B) bone marrow-derived macrophages from duplicate C57BL/6 mice were stimulated with 1 ng/ml IFN-γ for 30 minutes, with or without addition of 5 ng/ml IL-10. Cell extracts were prepared and assayed for STAT1, pSTAT1 (Y701), STAT3, and pSTAT3 (Y705). This experiment was performed twice and the figure shows the results of one representative experiment; (C) bone marrow-derived macrophages from three individual C57BL/6 mice were isolated and stimulated with 1 ng/ml IFN-γ and/or 5 ng/ml IL-10 for 2 hours. Cells were washed with phosphate-buffered saline and harvested into TRIzol reagent. RNA was isolated and the expression of Cxcl9, Cxcl10, and Cxcl11 were measured by quantitative reverse transcription-polymerase chain reaction. The figure shows the results from one representative experiment. GADPH = glyceraldehyde 3-phosphate dehydrogenase; GSEA = gene set enrichment analysis; IFN = interferon; IL = interleukin; STAT1 = signal transducer and activator of transcription 1; WT = wild-type.