Project description:This SuperSeries is composed of the following subset Series: GSE5099: Expression Data from Macrophage Maturation and Polarization Experiment GSE35433: Genome-wide analysis of human macrophages stimulated with IL-4 (20ng/ml) (Illumina) GSE35434: Genome-wide analysis of human macrophages stimulated with IL-4 (10ng/ml) (Illumina) GSE35435: Genome-wide analysis of mouse macrophages stimulated with IL-4 (bone marrow macrophages) (Affymetrix) GSE35436: Genome-wide analysis of mouse macrophages stimulated with IL-4 (Biogel and thioglycollate macrophages) (Affymetrix) Refer to individual Series
Project description:Bone marrow derived macrophages from C57BL/6 mice were stimulated into M1 and M2 polarization state. Analysis of BMDMs from LysMcre;FoxO1Fl/FL mice and control littermates. Results provide insight into the regulatory role of FoxO1 during macrophage polarization. BMDMs were stimulated with 100ng/ml LPS plus 20ng/ml IFN-γ into M1 polarization, and stimulated with 10ng/ml IL-4 plus 10ng/ml IL-13 into M2 polarization. Both for 24 hours. Unstimulated cells as M0 state.
Project description:Human aortic endothelial cells were stimulated by lysophosphatidylcholine (LPC) (10μM) with or without interleukin 35 (IL-35) (10ng/mL) or IL-10 (10ng/mL) for 18 hours. Total RNAs were extracted from samples, then mRNA and non-coding RNAs were enriched by removing rRNA from the total RNA. The library was sequenced by Illumina HiSeq4000 using PE100 strategy and the reads were mapped to the human hg19 reference genome.
Project description:Studying the effect of Th2 cytokine IL13 on on gene expression of human neutrophils by comparing gene expression of neutrophiles isolated from the blood of 3 healthy donors and stimulated with IL-13 (10ng/ml) or left unstimulated.
Project description:Hypoxia-inducible factor 1-alpha (HIF1a) is a basic helix-loop-helix PAS domain-containing protein, and is considered as the master transcriptional regulator of cellular response to hypoxia and inflammatory stimuli. Studies over the year have demonstrated that HIF1a play a critical role in innate immune cell response to infection and injury. Here we isolated bone marrow of 3 mice per group (Lyz2cre and Lyz2cre:HIF1aFl/Fl) and differentiated into macrophages by culturing them in M-CSF containing media. These cells were stimulated with 100ng/ml LPS, 10ng/ml IFNgamma, 10ng/ml IL4 or 10ng/ml IL10 for 18 hours. Total RNA was isolated and subjected to RNAseq analyses.
Project description:The cytokine IL-22 promotes tumor progression in murine models of colorectal cancer (CRC). However, the clinical significance of IL-22 in human CRC remained unclear. Using a rigorous discovery/verification analysis of tumor gene expression from over 1,000 patients, we discovered that among CRC patients with high expression of either or both subunits of the heterodimeric IL-22 receptor, KRAS mutation confers poor prognosis. Analysis of human CRC cell lines and primary tumor organoids, including a DLD-1 isogenic cell line pair that differed only in KRAS mutation status, showed that IL-22 and mutant KRAS cooperatively enhance cancer cell proliferation. The purpose of this study was to identify unique mechanisms of interaction between IL-22 signaling and mutant KRAS. Taking advantage of the DLD-1 isogenic pair with match IL-22 receptor expression, differing only in the presence or absence of mutant KRAS global transcriptomic changes were explored in an unbiased manner following IL-22 stimulation. RNA-sequencing was performed on DLD-1 isogenic cells stimulated with 10ng/mL IL-22 for 2h to identify early transcriptional changes and after 24h to explore late changes. Given that IL-6 receptor expression does not interact with mutant KRAS in a prognostically significant manner, nor did it enhance proliferation in KRAS mutant versus WT cells, DLD-1 isogenic cells were also stimulated with 10ng/mL IL-6 for 2 and 24h to identify pathways uniquely regulated by IL-22 and not IL-6.
Project description:Purpose: We found that IFN-g and IL-27 had suppressive effects on ILC2s cultured with IL-33. The goal of this study is to clarify the expressions of RNA induced by IFN-g and IL-27 in ILC2s. Methods: ILC2s were isolated from fat-asociated lymphid clusters (FALC) of wild-type mice. They were cultured with IL-33 (10ng/ml), IL-33 + IFN-g (10ng/ml), or IL-33 + IL-27 (10ng/ml) for 48hrs. RNA was isolated by Allprep DNA/RNA Micro Kit (QIAGEN), and cDNA libraries were prepared by TruSeq RNA Sample Preparation kits v2 (Illumina) according to the manufacturerâs low sample protocol. A HiSeq 1500 system (Illumina) was used for 50 single-end bases (50SE)ãsequencing. Results: Sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence, then mapped to the reference genome (mm9) using Bowtie2 v2.1.0 and TopHat2 v2.0.8. The transcript abundances were estimated as FPKM (fragments per kilobase of exon million fragments mapped) value using Cufflinks v2.1.1. We found that both IL-27 and IFN-g upregulated the expression of STAT1 and IRF1 which are regulated downstream of IFN-g receptor signaling, but there was no difference in the expression of GATA3, a critical transcription factor for ILC2 functions. Conclusions: Our study represents the detailed differences of RNA expressions by RNA-seq technology. RNA-Seq analysis of ILC2s cultured with IL-33 (10ng/ml), IL-33 + IFN-g (10ng/ml), or IL-33 + IL-27 (10ng/ml) for 48hrs.
Project description:The aim of the experiment is to identify chromatin and transcription factor binding dynamics in host gene expression upon infection of human Huh7 hepatoma carcinoma cells with human coronavirus HCoV-229E as compared to uninfected cells and in comparison to cells stimulated by IL-1alpha (10ng/ml) for 1h. All experiments have been performed at 33°C which is required for sufficient viral entry/replication.
Project description:Idiopathic pulmonary arterial hypertension (IPAH) is characterized by medial hypertrophy due to pulmonary arterial smooth muscle cell (paSMC) hyperplasia. Interleukin (IL)-13 is a potent regulator of tissue fibrosis and remodelling, and its effects are dependent on the cell-type specific expression of the IL-13 receptor isotypes IL-4Rα, IL-13Rα1, and IL-13Rα2. In order to identify the possible mechanism how IL-13 can exert its antiproliferative effect on paSMC microarray analysis was performed. For this purpose paSMC were stimulated with IL-13(10ng/ml) for 2h and 6h, respectively and subjected to microarray analysis.