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Protocol to quantify LRRK2-mediated Rab10 phosphorylation using high-content imaging. 使用高含量成像定量lrrk2介导的Rab10磷酸化的方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-04 DOI: 10.1016/j.xpro.2026.104819
Beatrice Masotti, Paolo Lorenzon, Giuseppina Covello, Elisa Greggio
{"title":"Protocol to quantify LRRK2-mediated Rab10 phosphorylation using high-content imaging.","authors":"Beatrice Masotti, Paolo Lorenzon, Giuseppina Covello, Elisa Greggio","doi":"10.1016/j.xpro.2026.104819","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104819","url":null,"abstract":"<p><p>Rab10 phosphorylation at Thr73 (pRab10) is a well-established readout of the kinase LRRK2, a protein whose mutations are associated with Parkinson's disease. Here, we present a protocol for high-content quantification of endogenous pRab10-positive vesicles in primary astrocyte-enriched cultures using the Operetta CLS system. We describe steps for cell culture, immunofluorescence, image acquisition, and quantitative analysis using Harmony software. This protocol is applicable to other LRRK2-expressing cells and offers a scalable platform for quantitative studies of LRRK2 physiological and pathological activity.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104819"},"PeriodicalIF":1.4,"publicationDate":"2026-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148892559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A concise guide to setting up a germ-free unit. 一个简明的指南，建立一个无菌的单位。
IF 1.4
STAR Protocols Pub Date : 2026-09-03 DOI: 10.1016/j.xpro.2026.104831
Romuald Binet, Claire F Pearson
{"title":"A concise guide to setting up a germ-free unit.","authors":"Romuald Binet, Claire F Pearson","doi":"10.1016/j.xpro.2026.104831","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104831","url":null,"abstract":"","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104831"},"PeriodicalIF":1.4,"publicationDate":"2026-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for optogenetic control of cAMP in human telencephalic organoids using an easy-to-build Arduino-driven LED system. 使用易于构建的arduino驱动LED系统对人类端脑类器官cAMP进行光遗传学控制的协议。
IF 1.4
STAR Protocols Pub Date : 2026-09-02 DOI: 10.1016/j.xpro.2026.104818
Issei S Shimada, Yoichi Kato
{"title":"Protocol for optogenetic control of cAMP in human telencephalic organoids using an easy-to-build Arduino-driven LED system.","authors":"Issei S Shimada, Yoichi Kato","doi":"10.1016/j.xpro.2026.104818","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104818","url":null,"abstract":"<p><p>Temporal regulation of cyclic adenosine monophosphate (cAMP) using photoactivated adenylyl cyclase (bPAC) is critical in research; however, commercial optogenetic systems are costly. Here, we present a protocol for optogenetic control of cAMP in human telencephalic organoids using an easy-to-build Arduino-driven light-emitting diode (LED) stimulation system. We describe steps for assembling the system, writing code for LED regulation, and preparing induced pluripotent stem (iPS) cells for optogenetic experiments. We then detail procedures for optogenetic activation of cAMP to control cell-fate decisions. For complete information on the generation and use of this protocol, please refer to Shimada et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104818"},"PeriodicalIF":1.4,"publicationDate":"2026-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148881915","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for laser-induced plasma membrane and cell wall damage in budding yeast. 激光诱导出芽酵母质膜和细胞壁损伤的方法。
IF 1.4
STAR Protocols Pub Date : 2026-09-02 DOI: 10.1016/j.xpro.2026.104814
Yuta Yamazaki, Keiko Kono
{"title":"Protocol for laser-induced plasma membrane and cell wall damage in budding yeast.","authors":"Yuta Yamazaki, Keiko Kono","doi":"10.1016/j.xpro.2026.104814","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104814","url":null,"abstract":"<p><p>Budding yeast is a model organism to study evolutionarily conserved biological processes, including plasma membrane (PM) repair. Here, we present a protocol to induce localized PM and cell wall damage during live-cell imaging. We describe steps for revival from frozen stock, liquid culture, inducing laser damage, and live-cell imaging. We then detail procedures for fluorescence-signal quantification and statistical analysis. This protocol enables the study of localized PM and cell wall repair mechanisms in budding yeast.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104814"},"PeriodicalIF":1.4,"publicationDate":"2026-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148881913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for modeling biliary cell activation in human precision-cut liver slices. 人类精确切割肝脏切片模拟胆道细胞活化的方法。
IF 1.4
STAR Protocols Pub Date : 2026-09-02 DOI: 10.1016/j.xpro.2026.104815
Beatrice Anfuso, Manuela Mastronardi, Silvia Palmisano, Giovanni Sorrentino
{"title":"Protocol for modeling biliary cell activation in human precision-cut liver slices.","authors":"Beatrice Anfuso, Manuela Mastronardi, Silvia Palmisano, Giovanni Sorrentino","doi":"10.1016/j.xpro.2026.104815","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104815","url":null,"abstract":"<p><p>Persistent ductular reaction is a hallmark of chronic liver injury, characterized by the proliferation and pro-inflammatory activity of reactive biliary epithelial cells. Here, we present a protocol for modeling biliary cell activation in human precision-cut liver slices. We describe steps for collecting samples, generating, and culturing precision-cut liver slices, and activating biliary cells ex vivo while preserving native architecture and multicellular interactions. Biliary cell activation can be quantified by EdU incorporation, supporting mechanistic studies and translational drug-screening applications. For complete details on the use and execution of this protocol, please refer to Anfuso et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104815"},"PeriodicalIF":1.4,"publicationDate":"2026-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148881882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for isolation of interstitial fluid from tumor-draining lymph nodes for proteomic profiling in murine breast cancer models. 从肿瘤引流淋巴结分离间质液用于小鼠乳腺癌模型蛋白质组学分析的方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-01 DOI: 10.1016/j.xpro.2026.104816
Greta Mattavelli, Jonathan J Swietlik, Moutaz Helal, Arpa Aintablian, Felix Meissner, Angela Riedel
{"title":"Protocol for isolation of interstitial fluid from tumor-draining lymph nodes for proteomic profiling in murine breast cancer models.","authors":"Greta Mattavelli, Jonathan J Swietlik, Moutaz Helal, Arpa Aintablian, Felix Meissner, Angela Riedel","doi":"10.1016/j.xpro.2026.104816","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104816","url":null,"abstract":"<p><p>Tumors release soluble factors that reach tumor-draining lymph nodes (TDLNs), where they induce remodeling already at the pre-metastatic stage and alter the local proteomic landscape. Here, we describe a protocol for isolating interstitial fluid from primary tumors and TDLNs in murine breast cancer models to study extracellular protein composition while preserving tissue viability and integrity. We outline key steps for tissue preparation, fluid extraction, and downstream bioinformatic analysis to support the characterization of tumor-associated changes in TDLNs. For complete details on the use and execution of this protocol, please refer to Mattavelli et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104816"},"PeriodicalIF":1.4,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148875158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Intraoral cannulation for controlled taste solution delivery in mice. 小鼠控制味觉溶液的口内插管。
IF 1.4
STAR Protocols Pub Date : 2026-09-01 DOI: 10.1016/j.xpro.2026.104811
Gyujin Park, Hojoon Lee
{"title":"Intraoral cannulation for controlled taste solution delivery in mice.","authors":"Gyujin Park, Hojoon Lee","doi":"10.1016/j.xpro.2026.104811","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104811","url":null,"abstract":"<p><p>Presentation of taste solutions is required for investigating the detection and processing of gustatory information. These stimuli are also widely used to motivate animals in behavioral tasks. Here, we present a protocol for surgically implanting an intraoral cannula in mice, enabling direct and precise delivery of fluids for passive stimulation of oral sensory systems (e.g., gustation, somatosensation, and nociception) and associated limbic pathways. We then detail procedures for controlled solution delivery. For complete details on the use and execution of this protocol, please refer to Park and Lee.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104811"},"PeriodicalIF":1.4,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148875125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for propagating and inoculating rice grassy stunt virus in rice plants using brown planthoppers. 利用褐飞虱在水稻植株上繁殖和接种水稻草矮病毒的方案。
IF 1.4
STAR Protocols Pub Date : 2026-08-31 DOI: 10.1016/j.xpro.2026.104813
Ming Wu, Guoyi Yang, Shanshan Zhao, Shuai Zhang, Jianguo Wu
{"title":"Protocol for propagating and inoculating rice grassy stunt virus in rice plants using brown planthoppers.","authors":"Ming Wu, Guoyi Yang, Shanshan Zhao, Shuai Zhang, Jianguo Wu","doi":"10.1016/j.xpro.2026.104813","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104813","url":null,"abstract":"<p><p>Rice grassy stunt virus (RGSV) causes a severe viral disease in rice and is transmitted by brown planthoppers (BPHs) (Nilaparvata lugens Stål). Here, we present a protocol for propagating RGSV and establishing controlled infection in rice plants. We describe steps for preparing rice seedlings, maintaining and propagating BPH colonies, generating viruliferous BPHs, and performing genotype-separated inoculation. This protocol provides a standardized framework for BPH-mediated RGSV inoculation and helps reduce vector-associated bias in infection assays across rice genotypes. For complete details on the use and execution of this protocol, please refer to Yang et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104813"},"PeriodicalIF":1.4,"publicationDate":"2026-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148865994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for haplotype-resolved structural variant detection via long-read sequencing using cuteHap. 利用cuteHap进行长读测序的单倍型解析结构变异检测方案。
IF 1.4
STAR Protocols Pub Date : 2026-08-28 DOI: 10.1016/j.xpro.2026.104810
Shuqi Cao, Chuanmin Wu, Yuejin He, Tao Jiang
{"title":"Protocol for haplotype-resolved structural variant detection via long-read sequencing using cuteHap.","authors":"Shuqi Cao, Chuanmin Wu, Yuejin He, Tao Jiang","doi":"10.1016/j.xpro.2026.104810","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104810","url":null,"abstract":"<p><p>Long-read sequencing technologies have revolutionized human genome exploration at an unparalleled resolution, particularly facilitating the analysis of structural variation (SV) at haplotype resolution. Here, we present a protocol for using cuteHap, a robust framework for haplotype-aware SV detection through phased alignment reads generated by diverse long-read sequencing platforms. We describe procedures for single-nucleotide variant (SNV) calling, read phasing, SV calling, and genotyping. We also establish a benchmarking pipeline to evaluate the detected SV callsets. For complete details on the use and execution of this protocol, please refer to Cao et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104810"},"PeriodicalIF":1.4,"publicationDate":"2026-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148857793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for model-guided alternating high-temperature thermotherapy to eliminate major viruses from diverse sugarcane cultivars. 模型引导的交替高温热疗法消除不同甘蔗品种主要病毒的方案。
IF 1.4
STAR Protocols Pub Date : 2026-08-27 DOI: 10.1016/j.xpro.2026.104808
Guo-Qiang Huang, Jin-Long Guo, Feng Li, Qian Dong, Tung-Yu Hsieh
{"title":"Protocol for model-guided alternating high-temperature thermotherapy to eliminate major viruses from diverse sugarcane cultivars.","authors":"Guo-Qiang Huang, Jin-Long Guo, Feng Li, Qian Dong, Tung-Yu Hsieh","doi":"10.1016/j.xpro.2026.104808","DOIUrl":"https://doi.org/10.1016/j.xpro.2026.104808","url":null,"abstract":"<p><p>Vegetatively propagated sugarcane can accumulate multiple viruses that compromise planting-material health. Here, we present a model-guided alternating high-temperature thermotherapy (AHTT) protocol to eliminate major viruses from diverse sugarcane cultivars. We describe preparation and disinfection of single-bud axillary-bud cuttings, planting, thermal preconditioning, and a 12-day AHTT regimen. We then detail post-thermotherapy recovery, health scoring, leaf sampling, total-RNA extraction, control-validated one-step RT-PCR, result interpretation, and the detection of additional sugarcane viruses. For complete details on the use and execution of this protocol, please refer to Huang et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 3","pages":"104808"},"PeriodicalIF":1.4,"publicationDate":"2026-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148841458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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