{"doi":"10.1016/j.xpro.2024.103400","title":"Isolation and transcriptional profiling of antigen-presenting cells from mouse lung and MLNs following intranasal immunization","abstract":"Dendritic cells (DCs) play a central role in the initiation of the adaptive immune response. Here, we present a protocol for isolating and transcriptionally profiling antigen-presenting cells (APCs) from the mouse lung and mediastinal lymph nodes (MLNs) following intranasal immunization. We describe steps for preparing single-cell suspensions from the lung and MLN, along with the detection and RNA sequencing (RNA-seq) of antigen-presenting DCs. This protocol offers a broadly applicable approach for identifying variations in DC subpopulations under diverse experimental conditions. For complete details on the use and execution of this protocol, please refer to Youhui et al. 1 • Steps for assessing antigen presentation by DCs in the mouse lung and MLN • Guidance on the preparation of single cells from the mouse lung and MLN • Steps for gating strategy for the identification of antigen-presenting DCs • Instructions for sorting DCs for transcriptome analysis Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Dendritic cells (DCs) play a central role in the initiation of the adaptive immune response. Here, we present a protocol for isolating and transcriptionally profiling antigen-presenting cells (APCs) from the mouse lung and mediastinal lymph nodes (MLNs) following intranasal immunization. We describe steps for preparing single-cell suspensions from the lung and MLN, along with the detection and RNA sequencing (RNA-seq) of antigen-presenting DCs. This protocol offers a broadly applicable approach for identifying variations in DC subpopulations under diverse experimental conditions.","journal":"STAR Protocols","year":2024,"id":481872,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.958,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1049887,"name":"Yihan Wang","orcid":"0000-0001-7440-417X","position":1,"is_corresponding":false},{"id":226717,"name":"Qiang Wang","orcid":"0000-0003-3123-3889","position":2,"is_corresponding":false},{"id":72297,"name":"Anita S. Chong","orcid":"0000-0003-0460-0196","position":3,"is_corresponding":false},{"id":524937,"name":"Shengbo Cao","orcid":"0000-0002-9787-7882","position":4,"is_corresponding":false},{"id":456814,"name":"Youhui Si","orcid":"0000-0001-8909-221X","position":5,"is_corresponding":false},{"id":1321806,"name":"Xiuyu Wang","orcid":"0000-0001-6293-988X","position":0,"is_corresponding":true}],"reference_count":8,"raw_metadata":null,"created_at":"2026-07-19T02:07:10.067421Z","pmid":"39427311","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}