GSM8586702: Control 3; Mus musculus; RNA-Seq
To isolate lung LECs, mice carrying the Prox1-EGFP reporter (in which all LECs are labeled with GFP) were injected with AlexaFluor647-conjugated isolectin (Invitrogen) for intravital labeling of endothelial cells just prior to sacrifice, as previously described. The lungs were then digested using dispase/collagenase in HBSS to generate a single cell suspension for FACS, with additional staining using antibodies for CD31 (Biolegend  102418), lineage cocktail (Bioloegend 133311), and Epcam (Biolegend 118225). LECs were sorted using positive selection for isolectin, CD31, GFP, and negative gating for Epcam and Lineage markers. 10X Genomics Chromium Single-cell 3' Library RNA-Seq Assays