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Next Generation Sequencing Facilitates Quantitative Analysis of Adipose Tregs from Foxp3Cre and Hif1afl/fl Foxp3Cre mice

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of Adipose Tregs from Foxp3Cre and Hif1afl/fl Foxp3Cre mice

PROVIDER: PRJNA638276 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11963179_1.fastq.gz	Fastqsanger.gz
	SRR11963179_2.fastq.gz	Fastqsanger.gz
	SRR11963180_1.fastq.gz	Fastqsanger.gz
	SRR11963180_2.fastq.gz	Fastqsanger.gz

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Next Generation Sequencing Facilitates Quantitative Analysis of Adipose Tregs from Foxp3Cre and Hif1afl/fl;Foxp3Cre mice

Project description:The goal of this study is to study mechanisms underlying how HIF-1α regulates adipose Treg function. Therefore, mRNA profiles of adipose Tregs (from 4-month old Foxp3Cre and Hif1afl/fl;Foxp3Cre mice) were generated by deep sequencing, single experiment, using Illumina HiSeq TEN. We compared gene expression profiles of adipose Tregs from Hif1afl/fl;Foxp3Cre and Foxp3Cre mice. Using unbiased comparative gene expression analyses, we found adipose HIF-1α-ΔTreg cells showed higher expression of Nt5e gene and simultaneously down-regulated canonical adipose Treg genes (including Pparg, Il1rl1, Klrg1, Areg). Therefore, these data suggested a dependency of HIF-1α on PPAR-γ expression in adipose Tregs.

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Project description:The goal of this study is to reveal unique features of adipose Tregs and to study mechanisms underlying how mediator Med23 regulate adipose Treg function in aged mice. Therefore, mRNA profiles of LN Tregs (from 4-month old Foxp3Cre mice) and adipose Tregs (from 4-month old Foxp3Cre, 4-month old obese Foxp3Cre, 10-month old Foxp3Cre and 10-month old Med23fl/fl;Foxp3Cre mice) were generated by deep sequencing, single experiment, using Illumina HiSeq2000. We compared transcriptome features between lymph node-derived Tregs and adipose Tregs from 4-month old lean or obese mice. Using unbiased comparative gene expression analyses, we found adipose Tregs display an up-signature of Insr (Insulin receptor) and Hif1a, while Pparg acts as a positive control. We next compared gene expression profiles of adipose Tregs from 10-month old Med23fl/fl;Foxp3Cre (MKO) and Foxp3Cre (WT) mice. adipose Med23-ΔTreg cells display impaired transcription of Pparg and Il1rl1 (ST2), and they simultaneously acquire the expression of Nt5e (CD73), while Entpd1 (CD39) expression was not dramatically altered. Our studies implicate protective roles of CD73hi adipose Tregs and offer new therapeutic strategies against age-associated metabolic syndrome.

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Gene expression profile of regulatory T cells (Tregs) isolated from visceral adipose tissue and lymph nodes of mice sufficient and deficient of Pparg expression in Tregs

Project description:We identified Pparg as a major orchestrator of the phenotype of adipose-tissue resident regulatory T cells (VAT Tregs). To establish the role of Pparg in shaping the VAT Tregs gene profile and cell dynamics, Tregs from lymph nodes and visceral adipose tissue of mice sufficient and deficient of Pparg expression in Tregs were double sorted for microarray analysis. All gene expression profiles were obtained from highly purified (double-sorted) T cell populations sorted by flow cytometry. To reduce variability, cells from multiple mice were pooled. Triplicates were generated for all groups. Raw data were preprocessed with the RMA algorithm in GenePattern and averaged expression values were used for analysis.

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Gene expression of visceral adipose regulatory T cells from mice with specific deletion of insulin receptor in the cells

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