Project description:To investigate the impact of FoxP3 multimerization mutants on FoxP3's genome-wide gene expression profiles, we conducted FoxP3 RNA-seq experiments using mouse CD4+ T cells. The results indicated that all of these multimerization mutants had an effect on FoxP3 gene expression profiles, including some well-established Treg signatures.
Project description:In this study, we compared the proteomes of mouse CD4+Foxp3+ regulatory T cells (Treg) and CD4+Foxp3- conventional T cells (Tconv) in order to build a data set of proteins differentially regulated in these two cell populations. The data set contains mass spectrometry results from the analysis of 7 biological replicates of Treg/Tconv cell samples purified by flow cytometry, each experiment performed from a pool of 4-5 mice. Global proteomic analysis of each sample was performed by single-run nanoLC-MS/MS, using chromatographic separation of peptides on 50cm C18 reverse-phase columns, with either a 480min gradient on LTQ-Velos orbitrap mass spectrometer (replicates 1 and 2) or a 300min gradient on Q-Exactive orbitrap mass spectrometer (replicates 3-7). Several MS injection replicates were performed for some experiments, leading to 27 raw files composing the data set. The detailed description of each analysis (file name, sample type, biological replicate number, MS technical replicate number, MS instrument used, sample name in MaxQuant ouput) is given in the table “Files list.txt”.
Project description:To investigate the impact of FoxP3 multimerization mutants on FoxP3's genome-wide DNA binding, with a particular focus on TnG-repeats binding, we conducted FoxP3 ChIP-seq experiments using mouse CD4+ T cells. The results revealed that all of these multimerization mutants have the potential to influence FoxP3 DNA binding, including its interaction with TnG-repeats.
Project description:Single cell transcriptomic analysis (InDrops) of mouse FoxP3- CD4+ Tconv cells and FoxP3+ regulatory CD4+ T cells isolated from spleen and colon
Project description:Single cell transcriptomic analysis (InDrops) of mouse FoxP3- CD4+ Tconv cells and FoxP3+ regulatory CD4+ T cells isolated from spleen and visceral adipose tissue
Project description:Single-cell RNAseq (10x Genomics) analysis of mouse splenic CD4+ T cells in WT and ∆Foxp3 mice and in WT/∆Foxp3 bone marrow chimeras. Mouse CD4+T cells in 21-day-old male WT and ∆Foxp3 mice were isolated from spleen by flow cytometry as DAPI–TCRβ+CD4+ cells for 10x Genomics Single Cell 3′ Reagent Kit (V2 chemistry, one sample per channel). For bone marrow chimera experiment: 7 week-old CD45.2-recipient mice were irradiated with 1000 Rad, reconstituted with 4 million CD3-depleted bone marrow cells: 50% CD45.1 x Foxp3-IRES-GFP (WT, 21d-old male) and 50% Foxp3DeltaEGFPiCre/RFP x ROSA-YFP x CD45.1/2 (scurfy, 21d-old male). 10 weeks later, spleen were harvested and tagged using a different Hashtags fo each mouse. ∆Foxp3 CD4+ T cells were sorted as DAPI–TCRb+CD4+CD45.1+CD45.2+. WT CD4+ T cells were sorted as DAPI–TCRb+CD4+CD45.1+CD45.2–. Control WT DAPI–TCRb+CD4+GFP+ Treg cells and GFP- Tconvs cells were also tagged and sorted. Samples with different hastags were pooled before single cell encapsulation using 10x Genomics Single Cell 3′ Reagent Kit (V3 chemistry).
