Protocol for transcriptomic and epigenomic analyses of tip-like endothelial cells using scRNA-seq and ChIP-seq.

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS STAR Protocols Pub Date : 2025-01-10 DOI:10.1016/j.xpro.2024.103326
Shintaro Funasaki, Yuri Miyamura, Shunsuke Kamei, Akhinur Rahman, Masaya Yamazaki, Shingo Usuki, Keiichiro Yasunaga, Yorifumi Satou, Hiroto Ohguchi, Takashi Minami
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Abstract

Angiogenesis begins as endothelial cells migrate, forming a sprouting tip and subsequent growth-rich stalk cells. Here, we present a protocol for transcriptomic and epigenomic analyses of tip-like cells in cultured endothelial cells. We describe steps for stimulating human umbilical vein endothelial cells (HUVECs) with vascular endothelial cell growth factor (VEGF) to generate tip-like cells. We then detail procedures for library preparation for single-cell RNA sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq), and data analysis. This scalable protocol is also applicable to diverse omics studies, including proteomics and metabolomics. For complete details on the use and execution of this protocol, please refer to Miyamura et al.1.

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使用scRNA-seq和ChIP-seq对尖端样内皮细胞进行转录组学和表观基因组学分析的方案。
血管生成开始于内皮细胞的迁移,形成芽尖和随后的生长丰富的茎细胞。在这里,我们提出了一种在培养的内皮细胞中对尖端样细胞进行转录组学和表观基因组分析的方案。我们描述了用血管内皮细胞生长因子(VEGF)刺激人脐静脉内皮细胞(HUVECs)产生尖端样细胞的步骤。然后详细介绍了单细胞RNA测序(RNA-seq)和染色质免疫沉淀测序(ChIP-seq)文库制备的程序,以及数据分析。这种可扩展的协议也适用于多种组学研究,包括蛋白质组学和代谢组学。有关本协议使用和执行的完整细节,请参考Miyamura等人1。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
STAR Protocols
STAR Protocols Biochemistry, Genetics and Molecular Biology-General Biochemistry, Genetics and Molecular Biology
CiteScore
2.00
自引率
0.00%
发文量
789
审稿时长
10 weeks
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