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Gene expression analysis on Naïve CD4 T cell with JMJD3 knockdown

ABSTRACT: Gene expression analysis on Naïve CD4 T cell with JMJD3 knockdown

PROVIDER: PRJNA495740 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8003420.fastq.gz	Fastqsanger.gz
	SRR8003421.fastq.gz	Fastqsanger.gz
	SRR8003422.fastq.gz	Fastqsanger.gz
	SRR8003413.fastq.gz	Fastqsanger.gz
	SRR8003423.fastq.gz	Fastqsanger.gz

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Gene expression analysis on Naïve CD4 T cell with JMJD3 knockdown

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H3K27me3 and JMJD3 ChIP-seq during naïve CD4 T cell activation

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Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T cell differentiation

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Gene expression profiling in mature OT-II TCR transgenic CD4 SP thymocytes, either Jmjd3- and Utx-deficient or -sufficient.

Project description:The biological functions of histone demethylases Jmjd3 and Utx remain poorly understood. We assessed such functions in developing T cells, using conditional (CD4-Cre-mediated) gene disruption, by inactivating Kdm6a and Kdm6b, respectively encoding Utx and Jmjd3, in immature CD4+CD8+ thymocytes. We compared microarray gene expression in mature (Va2hi CD24lo) mutant and wild-type CD4+CD8- thymocytes carrying the OT-II TCR transgene. We show that Jmjd3 and Utx redundantly promote H3K27Me3 removal at, and expression of, a specific subset of genes involved in terminal thymocyte differentiation, especially S1pr1, encoding a sphingosine-phosphate receptor required for thymocyte egress. Mature (Va2hi CD24lo) CD4 thymocytes were sorted from freshly prepared single-cell suspensions OT-II TCR transgenic thymocytes deficient for Utx and Jmjd3 (dKO, CD4-Cre conditional deletion of floxed Kdm6a and Kdm6b alleles), and from Cre-negative controls (wild-type). Total RNA was extracted from sorted thymocytes using the RNeasy Plus Mini Kit (Qiagen) and processed for microarray analyses (Affymetrix Mouse Exon 1.0 ST array) at the NCI microarray facility, following the manufacturer’s recommendation. Data is generated from 3 replicates from each experiment.

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