RNA-seq of mouse fetal intestines comparing wildtype littermates with Notch inhibited mutants

Whole intestines were harvested from fetuses from timed pregnant matings for wildtype littermates and endothelial specific Notch inhibited mutants (Jax strain #032613). Fetal stages were confirmed according to the Theiler staging chart (https://www.emouseatlas.org/emap/ema/staging_criteria/staging_criteria.html). Whole intestines (from the common bile duct through the cecum) were collected at E16-E16.5. Male and female intestines from each stage were pooled and dissociated to single cells for single cell RNA sequencing as previously described (Miller et al. 2020 Dev Cell). 705-KW-1 and 705-KW-2 were whole intestine. 2482-KW1 and 2482-KW2 samples were enriched with CD31+ MACS beads. 3593-KW1 and 3593-KW2 samples were enriched with CD31+ MACS beads and depleted for CD45.本研究从经定时交配的孕鼠胚胎中采集完整肠道组织，受试胚胎分为野生型同窝仔鼠与内皮特异性Notch信号抑制突变体（Jax品系#032613）两组。胚胎发育阶段依据泰勒分期表进行确认，具体链接为https://www.emouseatlas.org/emap/ema/staging_criteria/staging_criteria.html。采集的肠道为胆总管至盲肠段的完整肠道，采集时间为胚胎发育第16天至16.5天（E16-E16.5）。将各发育阶段的雌雄个体肠道组织混合后解离为单细胞，用于单细胞RNA测序，实验方法参照Miller等人2020年发表于《Developmental Cell》的研究报道。其中，705-KW-1与705-KW-2为完整肠道样本；2482-KW1与2482-KW2样本采用CD31阳性MACS磁珠进行富集；3593-KW1与3593-KW2样本则先经CD31阳性MACS磁珠富集，再去除CD45阳性细胞。