Titlebook: Regulatory T-Cells; Methods and Protocol Masahiro Ono Book 2023 The Editor(s) (if applicable) and The Author(s), under exclusive license to

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Dendritic-Cells

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Erin Waters,Cayman Williams,Alan Kennedy,David M. Sansomcases, expected during the analysis and design phases. They allow verifying whether the implementation of each use case is in accordance with the corresponding specification. We also present in this paper a brief summary of the environment that we developed for supporting the proposed use case drive

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Analysis of FOXP3 DNA Methylation Patterns to Identify Functional FOXP3+ T-Cell Subpopulationsgene transcription. This protocol shows that bisulfite sequencing (BS) distinguishes methylated and unmethylated cytosines within DNA and reveals the methylation status of individual CpGs in cells within each population, identifying functionally different FOXP3. subpopulations.

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In Vitro Analysis of CTLA-4-Mediated Transendocytosis by Regulatory T Cellsly tagged ligands mixed with the expanded Tregs. This methodology can be applied to testing of patients carrying CTLA-4 mutations, providing a robust model to assess the degree of functional disruption.

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Depletion of Treg by the Diphtheria Toxin Systemis of Tregs in the skin and skin-draining lymph node (dLN) by flow cytometry. This procedure of selective depletion of cutaneous Tregs can be applicable to other tissues and cells, to allow investigation of the role of tissue-resident cells in mice.