Immune crosstalk between pancreas, gut and liver through lymph node sharing [scRNA-seq]

To investigate tissue of origin effects on migratory dendritic cell (DC) gene expression in shared lymph nodes we performed bulk RNAseq of DCs from each tissue (liver, pancreas, duodenum). We then performed single cell sequencing of the DCs within the draining lymph nodes. scRNAseq of FACs purified migratory dendritic cells from liver, pancreas and duodenum (n=4 each tissue, each tissue is 2 mice pooled) and single cell sequencing of YFP+ cells sorted from CD11c-YFP pancreatic-duodenal lymph nodes (n=3 each lymph node, each lymph node is 10 mice pooled).为探究组织起源对共享淋巴结内迁移性树突状细胞（migratory dendritic cell, DC）基因表达的影响，我们对肝脏、胰腺、十二指肠来源的树突状细胞开展了批量RNA测序（bulk RNAseq）。随后，我们针对引流淋巴结内的树突状细胞实施了单细胞测序相关实验：其一为对来自上述三种组织的经荧光激活细胞分选（fluorescence-activated cell sorting, FACS）纯化的迁移性树突状细胞的单细胞RNA测序（single cell RNA sequencing, scRNAseq），每组组织设置4个生物学重复，每个重复样本由2只小鼠的细胞混合制备；其二为对从CD11c-YFP小鼠的胰十二指肠淋巴结中分选得到的YFP阳性细胞的单细胞测序，每组淋巴结设置3个生物学重复，每个重复样本由10只小鼠的细胞混合制备。