RNA-seq of HUVEC ± shRNA knockdown of SENCR

To assess the transcriptome of HUVEC where the SENCR is knocked down. SENCR is a lncRNA we show stabilizes the endothelial cell adherens junction by binding CKAP4 and indirectly localizing CDH5 at the cell membrane. The RNA-seq study was carried out to ascertain whether the effects of SENCR could be attributed to a function associated with sponging endogenous microRNAs that target proteins of the adgherens junction. HUVEC were cultured in media and transfected with lentivirus containing a short-hairpin RNA to SENCR and a GFP tag. Cells were tracked in the dish until all cells exhibited GFP positivity which would inducate all cells expressed the shRNA SENCR in lentivirus. Sister plates of cells were transduced with a scambled shRNA with GFP and were similarly tracked for 100% GFP expression. At ~3 days post-transduction, all experimental and control replicates were harvested for total RNA isolation.