Figure 2.

IL-23-treated neutrophils display a distinct gene expression profile. Sorted primary neutrophils were cultured with LPS or IL-23 in vitro, and microarray analyses of gene expression profiles were performed. (a) A heatmap comparison of gene expression profiles identifies lineage-restricted genes in untreated-Neu, LPS-Neu and IL-23-Neu cells. Genes with predominant expression in one cell type (at least two-fold greater than any other cell type) are organized in the heatmap. (b) Venn diagram showing upregulated genes 1695+916+1362=3973 and downregulated genes 2505+1506+1438=5449 in LPS-Neu and IL-23-Neu compared with untreated neutrophils. Approximately 34% of upregulated genes and 26% of downregulated genes are specifically differentially expressed in IL-23-Neu compared with untreated neutrophils. (c) Expression profiling of related cytokines in untreated-Neu, LPS-Neu and IL-23-Neu cells. Genes with predominant expression in one cell type (at least two-fold change compared with other cell types) are organized in the heatmap. Colors represent genes greater than (red) or less than (blue) the second-highest expression in three cell types. (d) The expression levels of cytokines including iNOS, TNF-α, IL-6, IL-17A, IL-17F and IL-22 were determined by real-time PCR after the isolated neutrophils were induced with LPS and IL-23 for different times. (e) The concentrations of IL-17A, IL-17F and IL-22 in the culture medium of isolated neutrophils induced with LPS and IL-23 for different times were measured by ELISA assays. Data are expressed as the mean±s.d. (N=3–5), with one representative experiment from three independent experiments shown. *P<0.05, **P<0.01 and ***P<0.001 for comparisons between the indicated groups. Data were analyzed by the Mann–Whitney U-test or two-way ANOVA analysis using SPSS software.