scRNA-seq over multiple time points of heart development in WT C57Bl6/J embryos and in Tbx1 mutant mice (Tbx1 KO). scRNA-seq over multiple time points of heart development in WT C57Bl6/J embryos and in Tbx1 mutant mice (Tbx1 KO)

We performed scRNA-seq over multiple time points of heart development in WT C57Bl6/J embryos and in Tbx1 mutant mice (Tbx1 KO). We dissected primarily the cardiac region but also the regions dorsal to the heart and the pharyngeal arches to capture the progenitor cells that migrate into the heart and the neural crest cells. For time points E10.5 and E11.5, we primarily dissected the regions behind the heart and included less of the overall cardiac region. We included pharyngeal arches for all time points and, at E11.5, we excluded the first arch. For Tbx1 null embryos, we also generated scRNA-seq data for WT and mutant embryos from the same pool of dissociated cells used for scATAC-seq. Overall design: For all embryos, we micro-dissected the tissue and dissociated using TryplE. For all samples, multiple replicates for WT and KO were used and we include all replicates below.

本研究针对野生型（Wild Type, WT）C57Bl6/J小鼠胚胎与Tbx1基因敲除（Tbx1 KO）小鼠的心脏发育多个时间点，开展了单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）。实验中我们主要解剖心脏区域，同时采集心脏背侧区域与咽弓组织，以捕获迁移至心脏的祖细胞及神经嵴细胞。针对E10.5与E11.5两个发育时间点，我们主要取材心脏后方区域，整体心脏区域的占比相对更低。所有时间点的样本均纳入咽弓组织，但在E11.5时间点中我们排除了第一咽弓。针对Tbx1纯合敲除胚胎，我们还利用与单细胞转座酶可及性测序（single-cell assay for transposase-accessible chromatin sequencing, scATAC-seq）相同的细胞解离池，完成了野生型与突变型胚胎的scRNA-seq数据构建。整体实验设计如下：所有胚胎均经显微解剖获取组织，并使用TryplE试剂完成细胞解离；所有样本均设置多份野生型与敲除型生物学重复，下文将展示全部重复样本的相关数据。