Pluto Bioinformatics

GSE145145: Targets of EOMES in mouse CD4+ T cells

Bulk RNA sequencing

To gain mechanistic insights into how EOMES regulates CD4+ T-cell differentiation and function, we performed an RNA-seq analysis using Eomes-GFP reporter mice (Eomes+/GFP) to isolate GFP+ (EOMES+) and GFP- (EOMES-) CD4+ T-cells by FACS sorting.; We also sequenced RNA from Eomes-deficient GFP+ and GFP- CD4+ T-cells isolated from EomesT/GFP knock-out mice, in which one Eomes allele is disrupted by GFP insertion and the DNA-binding domain of the other allele is deleted in T cells by Lck-driven Cre recombinase.; Hence, we had two populations of GFP+ cells where the EOMES locus was transcribed, one with a transcriptionally active EOMES protein (Eomes+/GFP) and another one with no transcriptionally active EOMES (EomesT/GFP), plus the respective GFP-negative controls isolated from the same mice.; A homogeneous population of EOMES-expressing CD4+ T cells were obtained by transferring nave sorted CD25- CD45RBhigh CD4+ T-cells isolated from Eomes+/GFP reporter or EomesT/GFP knock-out donor animals into Rag2-/- mice.; After three weeks of adoptive transfer GFP+ and GFP- CD4+ T-cell populations from these mice were FACS-sorted for transcriptome analysis by RNA sequencing. SOURCE: Ekaterina LuparGrosschedl Max Planck Institute of Immunobiology and Epigenetics

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