Optimized protocol for dissociation of mouse intestinal epithelium to viable single-cells [0.03%]
优化的小鼠肠上皮细胞分离方案使其能够高效存活并单细胞化
Ni Zhao,Peiling Geng,Yong Ge
Ni Zhao
The functional profiling of intestinal epithelial cells (IECs), which comprise multiple heterogeneous populations, is crucial for understanding host interactions with dietary and microbial factors. Here, we present a protocol for dissociati...
Protocol for recording visual maps in the mouse superior colliculus and visual cortex with intrinsic optical imaging [0.03%]
小鼠上丘和视觉皮层固有光学成像记录视觉图谱的协议
Flora Boutet-Porretta,Josien Visser,Arthur Matthys et al.
Flora Boutet-Porretta et al.
Intrinsic optical imaging (IOI) is a powerful approach to record visual-functional maps across the mouse superior colliculus (SC) and primary visual cortex (V1). Here, we present a protocol to map retinotopy and orientation preference in th...
Protocol for validating computationally predicted splice-altering variants using full-length gene reporter assays [0.03%]
用于全长基因报告基因检测的计算预测的剪接变异验证方案
Maria E Lukes,Kostantin Kiianitsa,Anthony Widjaja et al.
Maria E Lukes et al.
Up to half of human disease-causing variants may disrupt RNA splicing, but their computationally predicted effects require experimental validation. Here, we present a protocol to test splice-altering variants using full-length gene reporter...
Protocol for precision cutting and short-term culture of lymph node tissue slices for modeling of innate immune responses ex vivo [0.03%]
体外模拟固有免疫反应的淋巴结组织片精确切割和短期培养方案
Joannah R Fergusson,Mark Coles,Calliope A Dendrou et al.
Joannah R Fergusson et al.
Secondary lymphoid organs are highly organized for the initiation of immune responses. Here, we present a protocol that enables this microanatomical structure to be preserved during short-term culture, allowing ex vivo modeling of human inn...
Protocol for stable cell line production to express muscle-type nicotinic receptor [0.03%]
一种用于表达肌肉型烟碱受体的稳定细胞系生产的方案协议
Anna Li,David B Sauer,Yin Yao Dong
Anna Li
The adult muscle-type nicotinic acetylcholine receptor (AChR) is essential for neuromuscular transmission but is difficult to produce due to the requirement for coordinated subunit assembly. Here, we present a protocol for generating doxycy...
Protocol for large-scale, high-yield, high-purity extracellular vesicle purification from human plasma [0.03%]
人血浆中大量高效纯化高质量外泌体的方案
Saba Zanganeh,Gabriel F Arias,Allaura S Cone et al.
Saba Zanganeh et al.
Extracellular vesicles (EVs) hold promise as biomarkers and drug delivery vehicles; however, their broader use is limited by the lack of effective and scalable purification methods. We present a protocol for purifying EVs from human plasma ...
Protocol to analyze steps of ribosome biogenesis in nuclear and cytosolic fractions of yeast cells [0.03%]
用于分析酵母细胞核组分和胞质组分中核糖体生物发生步骤的方案
Ramin Zadali,Ulrike Topf
Ramin Zadali
Cell fractionation enables the study of compartment-specific RNA processing events in organisms with complex cell walls. Here, we present a protocol for isolating nuclear and cytosolic fractions from Saccharomyces cerevisiae. We describe sp...
Protocol for immunofluorescence detection and quantitative analysis of pH-dependent transcriptional condensates [0.03%]
pH依赖性转录凝聚物的免疫荧光检测和定量分析规范
Zhongyang Wu,Krishnan Raghunathan,Zhe Zhong et al.
Zhongyang Wu et al.
Acidic pH regulates the assembly of transcriptional condensates containing BRD4 and MED1 in a variety of mouse and human cells. Here, we present a protocol to image and quantify BRD4 condensates in bone marrow-derived macrophages. We descri...
Protocol to measure protein half-life in cell culture using heavy water [0.03%]
利用重水测量细胞培养中蛋白质半衰期的方法
Lorena Alamillo,Alexander Black,Matthew C Juber et al.
Lorena Alamillo et al.
Protein turnover, comprising the continued synthesis and clearance of proteins, is required for protein homeostasis and cell survival. Here, we present a protocol to measure protein turnover half-life in cultured cells using D2O labeling. W...
Protocol to annotate dendritic cell maturation types in vivo making use of lipid nanoparticle-based approaches [0.03%]
基于脂质纳米颗粒的体内注释树突状细胞成熟类型的协议
Karo Van Lil,Janne Swinnen,Greta R Webb et al.
Karo Van Lil et al.
The current panels used to annotate mature dendritic cells (DCs) by flow cytometry do not suffice to deconvolute homeostatic from immunogenic mature DCs. Here, we present a protocol to classify DCs along a homeostatic/immunogenic spectrum m...