design: RNAseq of human aortic endothelial cells (TeloHAECs) under unidirectional flow (UF) and disturbed flow (DF).
library_construction_protocol: Cells were cultured in 6-well plates with EGM-2 medium supplemented with 4% dextran (Sigma-Aldrich, St. Louis, MO, 31392). Hemodynamic flows were applied to using a 1o tapered stainless steel motorised cone (UMD-17 (Arcus20 Technology, Livermore CA). The rotation of the cone captures healthy unidirectional flow (UF) seen in athero-protective distal internal carotid artery, while the disturbed flow (DF) mimics athero-susceptible human carotid artery. During flow application to cells, the flow device was placed at 37C with 5%CO2. TeloHAECs subjected to UF and DF were lysed and RNA was extracted as mentioned below. Two technical replicates for each condition were pooled together to obtain an experimental replicate. TeloHAECs are grown in EGM-2 growth media (Lonza).  TeloHAECs subjected to UF and DF were lysed, and total RNA was collected using a NucleoSpin RNA kit (Macherey-Nagel, #740955). Using a TruSEQ mRNA RNA-SEQ library preparation protocol (Illumina provided), strand-specific RNA-SEQ libraries were prepared. 