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Thymic tuft cell single-cell mRNA analysis

ABSTRACT: The goal of the study was to sequence mRNA from tuft cells (identified as CD11c-;CD45-;EpCAM+;IL-25+ using Flare25 reporter mice) in C57BL/6 and Aire-/- mice. The data were used to investigate thymic tuft cell heterogeneity with and without AIRE protein expression.

ORGANISM(S): Mus musculus

PROVIDER: GSE114895 | GEO | 2018/06/03

REPOSITORIES: GEO

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Intestinal and thymic tuft cell RNA-seq

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promiscuous gene expression in the mouse thymus

Project description:Gene expression in different thymic stromal cells and subsets thereof was analyzed in 6-12 week old wild type (C57BL/6) and Aire knock-out (mixed background) mice. Thymic stromal cells were purified by sequential enzymatic digestion (collagenase, collagenase/dispase and trypsin) followed by gradient centrifugation and FACS sorting. Sort criteria were as follows: dendritic cells (CD11c+, F4/80 -), macrophages (F4/80+, CD11c-), cTECs (CD45–/lo, CDR1/Ly51+, Ep-CAM+) and mTECs (CD45–/lo, CDR1/Ly51–, Ep-CAM+). mTECs of wild-type and Aire knock-out mice were further subdivided according to CD80 expression levels. For microarray analysis total RNA from thymic stromal cell samples of two independent experiments was pre-amplified and biotinylated by two rounds of cDNA synthesis and in vitro transcription. Fluorescence readings were evaluated by using Microarray Suite 5.0 software. Keywords = thymus Keywords = promiscuous gene expression Keywords = AIRE Keywords: ordered

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Transcription profiling of mouse thymic stromal cells from 6-12 week old wild type (C57BL/6) and Aire knock-outs (mixed background)

Project description:Gene expression in different thymic stromal cells and subsets thereof was analyzed in 6-12 week old wild type (C57BL/6) and Aire knock-out (mixed background) mice. Thymic stromal cells were purified by sequential enzymatic digestion (collagenase, collagenase/dispase and trypsin) followed by gradient centrifugation and FACS sorting. Sort criteria were as follows: dendritic cells (CD11c+, F4/80 -), macrophages (F4/80+, CD11c-), cTECs (CD45â/lo, CDR1/Ly51+, Ep-CAM+) and mTECs (CD45â/lo, CDR1/Ly51â, Ep-CAM+). mTECs of wild-type and Aire knock-out mice were further subdivided according to CD80 expression levels.,For microarray analysis total RNA from thymic stromal cell samples of two independent experiments was pre-amplified and biotinylated by two rounds of cDNA synthesis and in vitro transcription. Fluorescence readings were evaluated by using Microarray Suite 5.0 software.

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