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. 2018 Jun 22;9:1445. doi: 10.3389/fimmu.2018.01445

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Copyright © 2018 Zhu, Dong, Ji, Jiang, Leung, Liu, Ng, Tsang, Jiao, Lam and Wong.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Identification of IL-37b target genes through transcriptional profiling. Human eosinophils and BEAS-2B cells were cocultured with or without IL-37b (200 ng/ml) pre-treatment for 10 min followed by stimulation with PGN (10 µg/ml) for 20 h. Total RNA of eosinophils was extracted and transcriptome analysis was performed to identify IL-37b target genes. (A) Heatmap of DEG between IL-37b-treated PGN sample and untreated PGN sample is shown. (B) Gene ontology (GO) term enrichment analysis of downregulated DEG, *P < 0.05 indicates significantly enriched GO term after Benjamini–Hochberg correction. (C) Fold of change for downregulated DE genes was depicted on a log₂ scale heatmap, with a yellow-to-red gradient indicating downregulation to upregulation. (D) Real-time qPCR validation of RNA-seq result. The GAPDH housekeeping gene was used as the internal reference and the relative gene expression was calculated using 2^{−ΔCt (Ct, target gene-Ct, GAPDH)}. Results are shown as mean ± SD, n = 3 per group, *P < 0.05 when compared between the denoted groups. Abbreviations: DEG, differentially expressed genes; PGN, peptidoglycan.

Identification of IL-37b target genes through transcriptional profiling. Human eosinophils and BEAS-2B cells were cocultured with or without IL-37b (200 ng/ml) pre-treatment for 10 min followed by stimulation with PGN (10 µg/ml) for 20 h. Total RNA of eosinophils was extracted and transcriptome analysis was performed to identify IL-37b target genes. (A) Heatmap of DEG between IL-37b-treated PGN sample and untreated PGN sample is shown. (B) Gene ontology (GO) term enrichment analysis of downregulated DEG, *P < 0.05 indicates significantly enriched GO term after Benjamini–Hochberg correction. (C) Fold of change for downregulated DE genes was depicted on a log₂ scale heatmap, with a yellow-to-red gradient indicating downregulation to upregulation. (D) Real-time qPCR validation of RNA-seq result. The GAPDH housekeeping gene was used as the internal reference and the relative gene expression was calculated using 2^{−ΔCt (Ct, target gene-Ct, GAPDH)}. Results are shown as mean ± SD, n = 3 per group, *P < 0.05 when compared between the denoted groups. Abbreviations: DEG, differentially expressed genes; PGN, peptidoglycan.