The interaction between IL-33 and TRIM28 in the regulation of macrophage polarization in an ST2-independent manner

Description

Ovarian cancer is one of the most common gynecological cancers and is known to be an immunogenic tumor. The tumor microenvironment provides optimal condition for the growth of ovarian cancer. Macrophages display a highly functional plasticity in the tumor microenvironment, which can respond to various signals. Switching macrophages’ phenotype is a potential therapeutic strategy. In malignant epithelial ovarian tumors, Interleukin-33(IL-33) highly expressed for increasing tumor grade. Our previous studies showed that IL-33 promotes the polarization of M2 macrophages and tumor grows in vivo by reshaping macrophage metabolism through activating the suppressor of tumorigenicity 2(ST2)-dependent signaling pathway. RNA-seq was total RNA extracted from WT or ST2KO BMDMs stimulated with IL-4 (25 ng/ml) for 24 hours. ChIP-seq assays were performed with SimpleChIP® Plus Enzymatic Chromatin IP Kit with anti-IL-33 and normal goat IgG antibodies. The ChIP-enriched DNA from WT or ST2KO BMDMs was purified for analysis.

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RNA-seq BMDMs from mice were stimulated with IL-4 (25 ng/ml) for 24 hours. Total RNA was extracted from WT and ST2KO BMDMs, and quantified using Agilent 2100 Bioanalyzer(California, USA). RNA-seq was performed by Shanghai Personal Biotechnology Co., Ltd(Shanghai, China). Briefly, sequencing was performed by using the Illumina Hiseq platform, and reads were aligned to GRCm38 by Ensembl(http://www.ensembl.org/)using Bowtie2. Differential expression analysis was performed by using the DESeq package and expression variations with both p-value and q-value less than 0.05 were considered significant. Each group includes three mice. ChIP-seq analysis ChIP assays were performed with SimpleChIP® Plus Enzymatic Chromatin IP Kit(Cell Signaling Technology, Boston, Massachusetts, USA) with anti-IL-33(R&D Systems, Minneapolis, USA) and normal goat IgG(Abcam, Cambridge, MA, USA) antibodies. The ChIP-enriched DNA from WT or ST2KO BMDMs was purified for analysis. The sequencing reads were mapped to mouse reference genome GRCm38 assembly using Bowtie2. High-throughput sequencing was performed by Shanghai Outdo Biotech Co., Ltd.

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