{"doi":"10.1247/csf.25011","title":"A quantitative method to monitor STING degradation with dual-luciferase reporters","abstract":"Stimulator of interferon genes (STING) triggers the type I interferon and inflammatory responses against a variety of DNA pathogens, which is essential to limiting viral infection and replication. STING activates the downstream kinase TBK1 at the trans-Golgi network (TGN) and is degraded at lysosomes through a process called lysosomal microautophagy. Impaired STING targeting to lysosomes results in the prolonged inflammatory signal, which may be associated with a variety of neurodegenerative and autoinflammatory diseases. Thus, development of methods to quantify STING degradation helps understand the mechanism of lysosomal microautophagy and its related diseases. Here we report a quantitative method to monitor STING degradation with two luciferases, firefly luciferase (FLuc) and Nanoluciferase (NLuc). The expression plasmid is composed of FLuc, a P2A self-cleavage site, and NLuc-tagged STING. FLuc intensity reflects the total amount of translated protein, serving as an internal control, while NLuc intensity corresponds to the amount of STING. Comparison of the NLuc/FLuc ratios at different time points after STING stimulation revealed the kinetics of decay of STING levels in live cells. This method should provide a useful complement to western blotting and fluorescence-activated cell sorter (FACS) analysis presently used to monitor STING degradation.Key words: innate immunity, STING, membrane traffic, lysosomal degradation, luciferase.","journal":"Cell Structure and Function","year":2025,"id":540953,"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":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9523,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1066746,"name":"Kanako Sato","orcid":"0000-0003-4863-0004","position":1,"is_corresponding":false},{"id":1429342,"name":"Ayumi Shinojima","orcid":null,"position":2,"is_corresponding":false},{"id":1227907,"name":"Shogo Koide","orcid":null,"position":3,"is_corresponding":false},{"id":1429343,"name":"Ruri Shindo","orcid":null,"position":4,"is_corresponding":false},{"id":1429344,"name":"Kazune Hongo","orcid":null,"position":5,"is_corresponding":false},{"id":247649,"name":"Kojiro Mukai","orcid":"0000-0001-9806-910X","position":6,"is_corresponding":false},{"id":1227334,"name":"Yoshihiko Kuchitsu","orcid":"0000-0003-0422-8056","position":7,"is_corresponding":false},{"id":247652,"name":"Tomohiko Taguchi","orcid":"0000-0003-0464-6370","position":8,"is_corresponding":false},{"id":1429341,"name":"Tsumugi Shoji","orcid":null,"position":0,"is_corresponding":true}],"reference_count":46,"raw_metadata":null,"created_at":"2026-07-19T02:52:42.901627Z","pmid":"40254425","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":[]}