Metabolic maturation of neonatal hearts driven by γ-linolenic acid in maternal milk [RXR ChIP-seq]

The goal of this study was to compare the cistrome profile of RXR (ChIP-seq) of perinatal hearts of cardiomyocyte-specific RXRab-deficient and WT mice, in order to identify genomic regions which are directly controlled by TF RXRa and RXRb. P0 hearts from wild type (WT) and cardiomyocyte-specific RXRab-/- (edKO) newborns were collected and analyzed. Individual P0 hearts were considered as individual biological replicates. Prior to cardiac tissue homogenization, each heart was sequentially fixed with DSG and FA. Chromatin was extracted and then sheared using a Covaris sonicator. Immunoprecipitation was performed with the corresponding antibody and Dynabeads A. Input was obtained by keeping a 10% total chromatin.

本研究旨在基于染色质免疫共沉淀测序（ChIP-seq），对比心肌细胞特异性RXRab双敲除与野生型（WT）小鼠围产期心脏的RXR顺式调控组（cistrome）图谱，以鉴定直接受转录因子（TF）RXRα与RXRβ调控的基因组区域。本研究收集并分析了野生型（WT）及心肌细胞特异性RXRab-/-（edKO）新生小鼠的出生后第0天（P0）心脏样本，每单个P0心脏均视为独立的生物学重复样本。在进行心脏组织匀浆前，需对每颗心脏依次采用二琥珀酰亚胺戊二酸酯（DSG）与甲醛（FA）进行交联固定。提取染色质后，使用Covaris超声破碎仪完成染色质剪切。实验中使用对应特异性抗体与Dynabeads A磁珠开展免疫沉淀实验，取总染色质的10%作为Input对照样本。