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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression data for MACS-isolated primary splenic T cells

ABSTRACT: We are interested in unveiling potential gene expression alterations underlying the T cell-related phenotypes observed in DUSP4-/- mice. We used microarray to obtain a macroscopic views of gene profiles comparing freshly isolated splenic T cells from WT and DUSP4-/- mice Magnetic-assisted cell sorting isolating T cells to 90-95% purity from freshly prepared splenocytes, followed by immediate RNA extraction.

ORGANISM(S): Mus musculus

SUBMITTER: Ching-Yu Huang

PROVIDER: E-GEOD-75319 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Dual-Specificity Phosphatase 4 Regulates STAT5 Protein Stability and Helper T Cell Polarization.

Hsiao Wan-Yi WY Lin Yu-Chun YC Liao Fang-Hsuean FH Chan Yi-Chiao YC Huang Ching-Yu CY

PloS one 20151228 12

Immune responses are critically regulated by the functions of CD4 helper T cells. Based on their secreted cytokines, helper T cells are further categorized into different subsets like Treg or Th17 cells, which suppress or promote inflammatory responses, respectively. Signals from IL-2 activate the transcription factor STAT5 to promote Treg but suppress Th17 cell differentiation. Our previous results found that the deficiency of a dual-specificity phosphatase, DUSP4, induced STAT5 hyper-activatio ...[more]

PMID: 26710253

Similar Datasets

Gene expression data for MACS-isolated primary splenic T cells

Project description:We are interested in unveiling potential gene expression alterations underlying the T cell-related phenotypes observed in DUSP4-/- mice. We used microarray to obtain a macroscopic views of gene profiles comparing freshly isolated splenic T cells from WT and DUSP4-/- mice

2016-02-25 | GSE75319 | GEO

DUSP4 deficiency due to promoter hypermethylation drives oncogenic JNK signaling and tumor cell survival in diffuse large B-cell lymphoma (methylation profiling data)

Project description:The epigenetic dysregulation of tumor suppressor genes is a major driver of human carcinogenesis. We have combined genome-wide methylation analyses with functional screening to identify novel candidate tumor suppressor genes in diffuse large B-cell lymphoma (DLBCL). We find that the dual-specificity phosphatase DUSP4 is aberrantly silenced in nodal and extranodal DLBCL due to promoter hypermethylation; ectopic expression of wild type DUSP4, but not of a phosphatase-deficient mutant, dephosphorylates c-JUN N-terminal kinase (JNK) and induces apoptosis in DLBCL cells. JNK inhibition prevents DLBCL survival in vitro and in vivo, and synergizes strongly with inhibitors of chronic active B-cell receptor signaling. Our results provide a mechanistic basis for the clinical development of JNK inhibitors in DLBCL, alone or in synthetic lethal combinations. RNA-seq expression profiling data set related to this experiment was also deposited at ArrayExpress under accession number E-MTAB-2925: http://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-2925/

2015-03-25 | E-MTAB-2926 | biostudies-arrayexpress

mRNA sequencing of SKMEL28 cells

Project description:A RNA-seq analysis was performed to characterize the molecular intermediates responsible for the deleterious effects upon DUSP4 loss in SKMEL28 cells. Methods:SKMEL28 cells were transfected with siRNA against DUSP4 or non-targeting control with or without Trametinib (Tram) at 0.25nM. After 16 hours, cell were collected and RNAseq experiments were performed. Results: the expression of MITF and some of its target genes were significantly downregulated in the absence of DUSP4 and completely rescued by trametinib treatment. Conclusions: In mutant melanoma cells, the expression of MITF and its related target genes is subject to regulation by the DUSP4-ERK axis.

2022-04-22 | GSE181467 | GEO

Characterization of MACS isolated microglia cells (CD11b positive cells) in comparison to all non-CD11b positive cells (termed non-target cell fraction (NTCF)) with proteomics

Project description:Microglia isolation from adult mouse brains remains difficult in comparison to microglia isolation from brains from newborn mouse brains as connective tissue, extracellular matrix materials and thicker myelin sheaths substantially influence traditional isolation protocols used with newborn mouse brains. Since the introduction of magnetic activated cell sorting (MACS), it has been reported that it is possible to also obtain microglia cells from adult mouse brains with a very good purity. However, most studies only used flow cytometry and/or qPCR to determine microglia enrichment and enrichment of traditional microglia marker proteins. Therefore we wanted to characterize in an unbiased way the proteomes of with MACS isolated microglia cells (CD11b positive cells) in comparison to all non-CD11b positive cells (termed non-target cell fraction (NTCF)), which consist of primarily astrocytes, neurons and oligodendrocytes. Therefore, we isolated from three adult mouse brains via MACS CD11b positive cells as well as the remaining CD11b negative cells (NTCF) to characterize the proteome of both cell fractions. The obtained proteomes were compared to a dataset of proteomic signatures for all CNS cell types determined by Sharma et al., (2015, doi: 10.1038/nn.4160) to determine the amount of enrichment of microglia-specific proteins.

2024-03-22 | PXD031973 | Pride

Isolation and characterisation of human gingival margin-derived STRO-1/MACS(+) and MACS(-) cell populations.

Project description: Not available

| S-EPMC4817556 | biostudies-literature

Expression data from BIO-treated human epidermal keratinocytes

Project description:Human keratinocytes isolated from foreskin were cultured and treated with a GSK-3beta inhibitor, BIO. The effect of BIO was evaluated by the cell growth, clony formation and differentiating markers. We used microarrays to detail the global program of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process. Human primary keratinocytes were subjected for RNA extraction. The total RNAs were hybridized on Affymetrix microarrays. The cultured cells exhibit a homogous cell morphology at passage 1 to passage 5. The cells were treated with BIO at passage 3 or 4.

2011-02-24 | E-GEOD-27509 | biostudies-arrayexpress

Differential expression of microRNA in Muc2-/- mouse colonic epithelial cells

Project description:Intestinal epithelia are protected by a layer of mucin secreted by goblet cells against mechanical and chemical injuries, potent causes of inflammation, and the most abundant secreted intestinal mucin is encoded by the Muc2 gene. Genetic deletion of Muc2 causes intestinal inflammation in early stage and tumors after 3 months. The underlying mechanisms are not clear, but epigenetic alterations, particularly, up- and down-regulated microRNAs are involved in the malignant transformation from colitis to cancer. We used miRNA array to profile the differential expression of the miRNAs in Muc2-/- mouse colonic epithelial lin comparison with those in wild-type mice. Total RNA were extracted from mouse colonic epithelial cells and Muc2-/- and +/+, and the RNA were hybridized on Affymetrix miRNA microarray to determine the alterations of miRNAs during colitis development and its malignant transformation from colitis to cancer. To the end, we found miRNA were differential expressed in the Muc2-/- mice, among them 20 miRNAs were significantly downregulated and 71 miRNAs were significantly upregulated in Muc2-/- mice, in comparison with Muc2+/+ mice (change fold >2 or <0.5; T<0.01, p value< 0.05, q value< 0.05).

2014-05-31 | E-GEOD-56577 | biostudies-arrayexpress

Comparison of the DNA methylation profiles of CD14+ monocytes from human peripheral blood with DCs and MACs obtained by exposure with GM-CSF/IL-4 and GM-CSF, respectively, and with mature DCs and MACs after LPS exposure

Project description:Comparison of the DNA methylation profiles of CD14+ monocytes from human peripheral blood with derived dendritic cells (DCs) and macrophages (MACs) obtained by exposure with GM-CSF/IL-4 and GM-CSF, respectively, and with mature DCs and MACs after lipopolysaccharide (LPS) exposure The methylation profiles of bisulfite-modified DNA of human CD14+ monocytes were compared with derived immature dendritic cells (iDCs) and macrophages (iMACs) following GM-CSF/IL-4 and GM-CSF incubation, and then activation/maturation with lypopolysaccharyde (LPS) using the Infinium HumanMethylation450 BeadChips (Illumina, Inc., San Diego, CA,). This platform allows the interrogation of >485,000 methylation sites per sample at single-nucleotide resolution, and comprises an average of 17 CpG sites per gene in the 99% of RefSeq genes. 96% of CpG islands are covered, with additional coverage in CpG island shores and the regions flanking them. The samples were hybridized in the array following the manufacturerâs instructions. Total DNA isolated by standard procedures from CD14+ cells (total monocytes, MOs) corresponding to three sets of samples of monocytes (MOs), derived immature DCs and MACs (iDCS and iMACS) and activated/mature DCs and MACs following incubation with LPS (mDCS and mMACs)

2015-12-11 | E-GEOD-71837 | biostudies-arrayexpress

Identifying ChIP-seq enrichment using MACS.

Project description: Not available

| S-EPMC3868217 | biostudies-literature

PRAME and CTCFL-reactive TCRs for the treatment of ovarian cancer

Project description:Recurrent disease emerges in the majority of patients with ovarian cancer (OVCA). Adoptive T-cell therapies with T-cell receptors (TCRs) targeting tumor-associated antigens (TAAs) are considered promising solutions for less-immunogenic ‘cold’ ovarian tumors. In order to treat a broader patient population, more TCRs targeting peptides derived from different TAAs binding in various HLA class I molecules are essential. By performing a differential gene expression analysis using mRNA-seq datasets, PRAME, CTCFL and CLDN6 were selected as strictly tumor-specific TAAs, with high expression in ovarian cancer and at least 20-fold lower expression in all healthy tissues of risk. In primary OVCA patient samples and cell lines we confirmed expression and identified naturally expressed TAA-derived peptides in the HLA class I ligandome. Subsequently, high-avidity T-cell clones recognizing these peptides were isolated from the allo-HLA T-cell repertoire of healthy individuals. Three PRAME TCRs and one CTCFL TCR of the most promising T-cell clones were sequenced, and transferred to CD8+ T cells. The PRAME TCR-T cells demonstrated potent and specific antitumor reactivity in vitro and in vivo. The CTCFL TCR-T cells efficiently recognized primary patient-derived OVCA cells, and OVCA cell lines treated with demethylating agent 5-aza-2′-deoxycytidine (DAC). The identified PRAME and CTCFL TCRs are promising candidates for the treatment of patients with ovarian cancer, and are an essential addition to the currently used HLA-A*02:01 restricted PRAME TCRs. Our selection of differentially expressed genes, naturally expressed TAA peptides and potent TCRs can improve and broaden the use of T-cell therapies for patients with ovarian cancer or other PRAME or CTCFL expressing cancers.

2023-03-10 | PXD040651 | Pride

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