Protocol for identifying recirculating thymic regulatory T cells and characterizing the role of Eos in their function using scRNA-seq and TCR-seq.

Xuan Xie, Tom Hill, Francisco Otaizo-Carrasquero, Xunyong Zhou, Ethan M Shevach

Abstract read

In one paragraph

Article in STAR protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0cells of the map it votes in

0citing papers in PubMed

–field-weighted citation impact

1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

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3 · Its place in the literature

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No citing paper in PubMed yet.

4 · The record

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PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Xuan XieLaboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. Electronic address: xxsabrina0408@outlook.com.

Tom HillIntegrated Data Sciences Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.

Francisco Otaizo-CarrasqueroGenomic Research Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.

Xunyong ZhouBestzyme & Bestlife Group, Shuyang County, Suqian City, Jiangsu, China.

Ethan M ShevachLaboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. Electronic address: eshevach@niaid.nih.gov.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Previous studies on the characterization of regulatory T cells (Tregs) in the thymus have proven to be simplistic. Here, we present a protocol for identifying recirculating thymic Tregs in mice and characterizing the role of Eos in their function. We describe steps for thymic dissection, magnetic cell separator (autoMACS) and fluorescence-activated cell sorting (FACS), single-cell sequencing, and downstream computational analysis. This protocol details a refined classification based on analysis of gene expression from single-cell RNA sequencing (scRNA-seq) and TCR sequencing data. For details on the use of this protocol, refer to Xie et al.

Indexed as

Receptors, Antigen, T-CellRNA-SeqSequence Analysis, RNASingle-Cell Gene Expression AnalysisThymus GlandT-Lymphocytes, RegulatoryAnimalsFlow CytometryMiceReceptors, Antigen, T-CellBioinformaticsCell BiologyCell isolationDevelopmental biologyFlow CytometryGenomicsImmunologyModel OrganismsRNAseqSingle Cell

Identifiers

PMID42258356

DOI10.1016/j.xpro.2026.104620

PMCPMC13264222

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LicenceCC BY-NC-ND

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.