Transcriptome analysis of endothelial cells of lymphatic vessel subtypes. Transcriptome analysis of endothelial cells of lymphatic vessel subtypes

To identify genes regulating the functional specification of collecting lymphatic vessels, we performed transcriptome profiling of dermal ECs isolated from adult mouse ear skin by flow cytometry. Overall design: Dermal cell suspensions from ear skin of 5-week-old mice carrying a lymphatic endothelial cell (LEC) reporter Prox1-GFP (Tg(Prox1-EGFP)KY221Gsat) were first subjected to enrichment for ECs using PECAM1 antibody-coated magnetic beads, followed by sorting of live BEC and LEC populations based on the expression of the lymphatic markers PDPN and Prox1-GFP. LEC subpopulations were further defined by the level of LYVE1 expression, and assigned as LYVE1-high capillaries (cap LEC), LYVE1-low/- collecting vessels (col LEC) and LYVE1-intermed pre-collecting vessels (pre-col LEC). Blood endothelial cells (BEC) were sorted as PECAM1+ PDPN- Prox1-GFP- LYVE1- population. Three replicates were performed for each group.