The in vivo endothelial cell translatome is highly heterogeneous across vascular beds (HUVEC RNAseq)

The interspersed anatomic distribution of endothelial cells (ECs) and their dependence on microenvironmental cues have been major challenges in studying EC function and response to (patho)physiological stimuli in vivo. Combining EC-specific translating ribosome affinity purification (EC-TRAP) with RNA sequencing provides an accurate in vivo snapshot of tissue-specific EC expression profiles, and maintains a high degree of sensitivity to detect low abundant transcripts while limiting changes in gene expression profiles due to enzymatic tissue dissociation required to generate single cell suspensions for FACS sorting or single cell analysis. In vitro HUVEC data was obtained to ensure short-term cycloheximide treatment, as used for in vivo experiments, did not alter global gene expression. RNASeq data from freshly isolated (passage 1) or commercially available HUVECs (passage 10), incubated with 100µg/ml cycloheximide or 2µg/ml homoharringtonine for 5 minutes (n=4 per condition per passage).