RNA-seq of human brain and dermal microvascular endothelial cells treated with different doses of TNF for various time points compared to untreated controls
Samples were collected directly in the well using a lysing solution (1% 2-mercaptoethanol + Buffer RLT; RNeasy Mini Kit, 74106, Qiagen). Study 1: Human brain microvascular endothelial cells (HBMEC) and human dermal microvascular endothelial cells (HDMEC) were stimulated with either 0ng/mL, 1ng/mL or 10ng/mL of TNF for 0 hours, 10 minutes (0.167 hours), 30 minutes (0.5 hours), 2 hours, or 6 hours. Study 2: HBMEC were stimulated with either 1ng/mL or 10ng/mL of TNF for 0 hours, 6 hours, 12 hours, or 24 hours. RNeasy mini kit (Qiagen, 74106) buffers and protocol. RNA-sequencing libraries were prepared using the TruSeq Stranded mRNA kit (20020595, Illumina following the manufacturers protocol.