Protocol for chromatin accessibility profiling of human endothelial cells cultured under fluid shear stress using ATAC-seq

利用ATAC-seq技术对在流体剪切应力下培养的人类内皮细胞进行染色质可及性分析的方案

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作者:Lion Raaz ,Paul-Lennard Mendez ,Stefan Mundlos ,Petra Knaus ,Jerome Jatzlau

Abstract

Chromatin accessibility influences gene regulation and can be quantified using assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq). Recapitulating in vivo fluid shear stress (FSS) mechano-regimes in vitro allows the study of atheroprone and atheroprotective mechanisms. In this protocol, we show how to culture and harvest endothelial cells from microfluidic channels for the preparation of ATAC-seq, highlighting optional growth factor stimulation and different FSS rates. This extends the application of ATAC-seq to the analysis of in vitro mechanically stimulated cells. For complete details on the use and execution of this protocol, please refer to Jatzlau et al.1. Keywords: Cell Biology; Genomics; Molecular Biology.

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