Illumina HiSeq 2500 sequencing; Single cell RNA-sequencing of epithelial cells, endothelial cells and macrophages from mouse submandibular salivary glands
Salivary gland tissue was excised, enzymatically and mechanically digested and Epcam+ cells (epithelial cells), CD31+ cells (endothelial cells) and CD45+/CD11b+/F480+ cells (macrophages) were isolated using fluorescence-activated cell sorting (FACS) and mixed in the ratio 45%, 45%, 10%, respectively. Isolated cells were loaded into the Chromium Single Cell controller (10x Genomics). Nucleic acid extraction was performed following the manufacturer's instructions (10x Genomics, as part of the Chromium Single Cell 3' v3.1). The four libraries (one for each sample) were pooled for sequencing, taking into account the expected differences in cell numbers between each library.