Pan-AcH4 ChIP-chip with High Resolution Tiling Arrays on HUVECs and HuAoVSMCs

Differential EC enrichment of pan-AcH4 was assessed at 34 select genes, including 19 EC-enriched genes, 6 broadly expressed genes, and 9 EC-excluded genes, by pan-AcH4 (AcH4K5, AcH4K8, AcH4K12, and AcH4K16) ChIP-chip analysis on human umbilical vein endothelial cells (HUVECs) and human aortic vascular smooth muscle cells (HuAoVSMCs) with a high resolution tiling DNA microarray. Specifically, the tiling array analyzes the 50 kb genomic region upstream of transcription initiation, the intragenic regions , and 50 kb genomic region downstream of the last exon. Pan-AcH4 ChIP was conducted on HUVECs and HuAoVSMCs. These ChIP samples were hybridized to a custom 1 M agilent microarray for ChIP-chip analysis.

本研究借助高分辨率DNA平铺微阵列，以人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVECs）和人主动脉血管平滑肌细胞（human aortic vascular smooth muscle cells, HuAoVSMCs）为实验材料，通过泛乙酰化组蛋白H4（pan-AcH4）染色质免疫沉淀微阵列分析（ChIP-chip），对34个预选基因的泛AcH4在内皮细胞中的差异富集特征进行评估。本次研究涉及的基因共分为三类：19个内皮细胞富集基因、6个广泛表达基因与9个内皮细胞排除基因，其中泛AcH4包含AcH4K5、AcH4K8、AcH4K12及AcH4K16四种修饰亚型。
具体而言，该平铺微阵列的检测区间涵盖转录起始位点上游50 kb的基因组区域、基因内部区域，以及最后一个外显子下游50 kb的基因组区域。
本研究首先针对HUVECs与HuAoVSMCs开展泛AcH4的染色质免疫沉淀（ChIP）实验，随后将获取的ChIP样本与定制1兆探针安捷伦（Agilent）微阵列进行杂交，以此完成ChIP-chip分析。