Illumina HiSeq 4000 paired end sequencing; Gene expression data in control (shCtrl) and B55alpha-silenced (shB55a KD) HUVECs
Gene expression data in control (shCtrl) and B55alpha-silenced (shB55a KD) HUVECs
HUVECs cultured in 6-well plates were washed in PBS once and then harvested in 1ml Trizol. HUVECs were isolated from human umbilical cords and cultured in Gelatin coated cell culture dishes, using M199 Medium supplemented with 20% FBS, PenStrep, Glutamine, heparin and endothelial cell growth factors (ECGS, PromoCell).  300000 HUVECs (passage 6) were seeded in 2ml of medium per well of a 6-well plate, treated with lentiviral shCtrl and shB55alpha (Sigma; TRCN0000002489) and cultured in the presence of qVD for 20hrs in Full Medium.  RNA was isolated using Trizol and the concentration as well as purity was determined spectrophotometrically using the Nanodrop ND-1000 (Nanodrop Technologies). Per sample, an amount of 3000ng of total RNA was used as input. Subsequently, preparation was performed with KAPA stranded mRNA seq Kit according to the protocol and samples were pooled equimolar.  Preparation was performed with KAPA stranded mRNA seq Kit according to the protocol and samples were pooled equimolar.
Experimental Factor: RNA interference
B55alpha knockdown by siRNA