Protocol for in vitro chemoenzymatic synthesis of thiazole-containing macrocyclic peptides via ribosomal incorporation of thioamides [0.03%]
通过核糖体翻译硫胺类非蛋白氨基酸实现含有噻唑环的巨环肽的体外化学酶法合成protocols
Akihiro Saito,Yuki Goto
Akihiro Saito
Backbone thiazole moieties (Thz) are prevalent structural features in peptidic natural products and contribute to favorable physicochemical properties. Here, we describe a protocol for in vitro chemoenzymatic synthesis of artificial Thz-con...
Protocol for a primary human 2D intestinal epithelium-dendritic cell co-culture organoid model to study mucosal immunity and viral infections [0.03%]
人源性二维肠上皮细胞-树突状细胞共培养类器官模型协议，用于研究黏膜免疫和病毒感染问题
Anusca Germaine Rader,Carla Mónica Sampaio Ribeiro
Anusca Germaine Rader
The intestinal mucosa is a major HIV-1 infection site, making immune-augmented intestinal models essential for mechanistic and therapeutic studies. We present a protocol that converts human intestinal organoids into polarized 2D epithelial ...
Protocol for cell-specific ratiometric quantification of apoplastic pH in Arabidopsis seedlings [0.03%]
拟南芥幼苗细胞特异性液泡胞外pH比值定量的协议
Lu Xiong,Paloma Mas
Lu Xiong
Apoplastic pH dynamically regulates plant intercellular communication, but its measurement in internal tissues, such as the vasculature, remains technically challenging. Here, we present a protocol for ratiometric quantification of apoplast...
Protocol for high-resolution multiplexed sequencing of single-cell full-length transcriptome with CBTi-seq [0.03%]
基于CBTi-seq的单细胞全长转录组高通量测序protocol
Liyong He,Wenyi Zhang,Peidong Qi et al.
Liyong He et al.
Balancing throughput and full-length coverage remains challenging in single-cell RNA sequencing (scRNA-seq). Here, we present combinational barcoded Tn5 transposon insertion sequencing (CBTi-seq), a highly scalable protocol for constructing...
Protocol for automated optimization of feature selection and clustering parameters in single-cell RNA-seq using scAutoTune [0.03%]
使用scAutoTune自动优化单细胞RNA测序中特征选择和聚类参数的协议
Shi Hai Sun,John S Pezaris
Shi Hai Sun
Single-cell RNA sequencing (RNA-seq) analysis requires dataset-specific tuning of feature selection and clustering parameters. Here, we present a protocol for automated optimization of feature selection and clustering parameters in single-c...
Protocol for single-cell epigenetic profiling in human organoids and tumoroids with Epi-CyTOF [0.03%]
Epi-CyTOF人器官体和肿瘤类器官单细胞表观遗传谱分析方案
Ezgi Senoglu,Vivian Mittné,Sevina Dietz et al.
Ezgi Senoglu et al.
In this protocol, we present Epi-CyTOF, a cytometry by time of flight (CyTOF) approach for single-cell epigenome profiling in cortical organoids as a model system for human brain development. We describe steps for in silico panel design, an...
Protocol for the enrichment of endosteal and periosteal mesenchymal cells from murine bone for single-cell transcriptome analysis [0.03%]
小鼠骨内皮细胞和骨膜细胞富集的协议及其单细胞转录组分析
Alongkorn Kurilung,Visanu Wanchai,Jinhu Xiong et al.
Alongkorn Kurilung et al.
Bone comprises diverse cell populations essential for skeletal development, remodeling, and homeostasis. Current single-cell isolation methods often use bulk digestion that can obscure distinctions between periosteal and endosteal compartme...
Protocol for PI3K inhibitor-free differentiation and maturation of human iPSC-derived arterial- and venous-like endothelial cells [0.03%]
人诱导多能干细胞来源的动脉和静脉类似内皮细胞的分化成熟无需PI3K抑制剂的方案
Oliwia N Mruk,Alexandra J Musk,Ralitsa R Madsen
Oliwia N Mruk
Vascular malformations caused by genetic PI3K pathway activation are highly debilitating and hard to treat. Here, we present a PI3K inhibitor-free protocol to differentiate arterial and venous endothelial cells from human induced pluripoten...
Protocol to stress human primary pancreatic islets with diabetogenic factors for subsequent single-cell RNA sequencing [0.03%]
用于糖尿病因子诱导压力的原代胰岛细胞单细胞RNA测序协议
Daniel A Veronese-Paniagua,Marlie M Maestas,Jared P Taylor et al.
Daniel A Veronese-Paniagua et al.
Type 1 diabetes (T1D) is an autoimmune disease that destroys insulin-secreting pancreatic β cells. Using human islets is instrumental to understanding the initiation and progression of T1D. Here, we present a protocol for treating cadaveri...
Protocol for multiplexed quantification of fluorescent biosensor FRET efficiency using barcoded cells [0.03%]
条形码细胞荧光生物传感器FRET效率多重定量方法学
Jhen-Wei Wu,Yichu Xu,Yun Chen et al.
Jhen-Wei Wu et al.
Genetically encoded fluorescent biosensors frequently use Förster resonance energy transfer (FRET) between donor and acceptor fluorescent proteins (FPs) as readouts for monitoring molecular activities in live cells. Here, we present a prot...