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. 2018 Mar 5;15(8):782–793. doi: 10.1038/cmi.2017.163

Figure 2.

Figure 2

IL-33-treated neutrophils express distinct gene profiles different from the resting N0 or N(LPS) inflammatory subpopulation. Sorted primary neutrophils were cultured with LPS or IL-33 in vitro and microarray analysis of gene expression profiles were performed. (a) Heatmap comparison of gene expression profiles identifies lineage-restricted genes in N0, N(LPS) and N(IL-33) cells. Genes with predominant expression levels in one cell type (at least twofold greater than any other cell types) were organized to draw the plot. (b) Venn diagram showing upregulated genes (n=3 905) and downregulated genes (n=6 497) in N(LPS) and N(IL-33) neutrophils compared with N0 neutrophils. About 33% of upregulated genes and 38% of downregulated genes were specifically differentially expressed in N(IL-33) compared with N0 cells. (c) Expression profiling of lineage-restricted cytokines and chemokines in N0, N(LPS) and N(IL-33) cells. Genes with predominant expression levels in one cell type (at least twofold change compared with other cell types) were organized to draw the plot. Colors represent genes above (red) or below (blue) the second highest expression in three cell types. (d) The levels of cytokines, including TNF-α, IL-1β, IL-4, IL-5, IL-9 and IL-13 expression levels, were determined by real-time PCR after the isolated neutrophils were induced with LPS and IL-33 for the indicated times. (e) The concentrations of TNF-α, IL-1β, IL-9, IL-5 and IL-13 in the culture medium of the isolated neutrophils induced with LPS and IL-33 for indicated times were detected by ELISA assays. (f) The levels of chemokines, including CXCL1, CCL5, CCL2, CXCL3, CCL7 and CCL12, were determined by real-time PCR. Data are expressed as mean±s.d. (n=3–5), showing one representative of the three independent experiments. *P<0.05, **P<0.01 and ***P<0.001 compared with the control or LPS-treated cells. CCL, C-C motif chemokine ligand; CXCL, C-X-C motif chemokine ligand; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; LPS, lipopolysaccharide; TNF, tumor necrosis factor.