Shear stress-induced long non-coding RNA, NIMBUSS, is required for the KLF2-mediated gene expression profile associated with vascular homeostasis I

The endothelial cell (EC) response to shear stress is partly mediated by the transcription factor KLF2, which is preferentially activated by laminar flow and promotes an atheroprotective phenotype. The up-regulation of KLF2 elicits anti-inflammmatory, anti-coagulant, and pro-vasodilatory processes. To investigate the function of the flow-induced long non-coding RNA (lncRNA) NIMBUSS, we performed loss-of-function studies using siRNA for NIMBUSS in ECs overexpressing KLF2. Four independent human umbilical vein endothelial cells (HUVEC) lines were transduced with adenovirus overexpressing GFP only or murine KLF2, and transfected with either control or two independent NIMBUSS siRNAs (16 samples: 8 Ad-GFP, 8 Ad-KLF2). Total RNA was isolated and analyzed using the Human lncRNA Microarray V3.0 (Arraystar) which profiles the expression of 30,586 lncRNAs and 26,109 protein-coding genes.