Cut&Tag profiling of NR1D1 in primary mouse vascular smooth muscle cells with or without vasostatin-2 stimulation

NR1D1 (also known as Rev-erba) is a nuclear receptor involved in circadian rhythm and metabolic regulation. Its role in vascular biology is not fully understood. We performed CUT&Tag (Cleavage Under Targets and Tagmentation) to investigate the genome-wide binding profile of the transcription factor NR1D1 in primary vascular smooth muscle cells (VSMCs) isolated from mice. Two experimental conditions were included: cells treated with vasostatin-2 and untreated controls. The goal of this study was to identify potential transcriptional regulatory targets of NR1D1 under basal and vasostatin-2-stimulated conditions. CUT&Tag was carried out using a specific antibody against NR1D1, and sequencing was performed on the Illumina NovaSeq 6000 platform. Overall design: CUT&Tag was performed using an NR1D1 antibody in primary mouse vascular smooth muscle cells with or without vasostatin-2 treatment to investigate differential chromatin binding under stimulated and control conditions.

NR1D1（亦称Rev-erba）是一类参与昼夜节律调控与代谢调控的核受体，其在血管生物学领域的功能尚未完全阐明。本研究借助CUT&Tag（靶向切割与标签化，Cleavage Under Targets and Tagmentation）技术，对从小鼠体内分离的原代血管平滑肌细胞（VSMCs）中转录因子NR1D1的全基因组结合谱展开探究。实验设置两组处理条件：经血管抑素-2刺激的细胞组，以及未接受处理的空白对照组。本研究的核心目标为鉴定NR1D1在基础稳态与血管抑素-2刺激条件下的潜在转录调控靶标。实验过程中，使用针对NR1D1的特异性抗体完成CUT&Tag实验，并在因美纳NovaSeq 6000测序平台上完成高通量测序。整体实验设计如下：在经或未经血管抑素-2处理的原代小鼠血管平滑肌细胞中，采用NR1D1抗体开展CUT&Tag实验，以解析刺激组与对照组间的染色质结合差异。