title: NextSeq 500 sequencing; Transcriptome analysis of human brain microvascular endothelial cells response to interactive DIII domain of protein E of WNV and interactive DIII domain of protein E of TBEV using RNA-seq
design: Transcriptome analysis of human brain microvascular endothelial cells response to interactive DIII domain of protein E of WNV and interactive DIII domain of protein E of TBEV using RNA-seq
library_construction_protocol: Human brain microvascular endothelial cells (hBMECs) were cultured in 6-well plates then incubated either with recombinant DIII of protein E of WNV or recombinant DIII of protein E of TBEV or culture medium only (non-induced control) for 6 hours.  Human BMEC (hBMEC/D3 cell line), was acquired from Merck/Millipore (Prague, Czech Republic). Cells were cultured in 6-well cell culture plates covered with collagen type I (Sigma, USA) and endothelial cell growth basal medium (EBM-2; Lonza,UK; Appendix 1). Cells were incubated at 37°C in a humid atmosphere in the presence of 5% CO2.    Human brain endothelial cells cultured in 6-well plates were incubated either with recombinant DIII of protein E of WNV (27 μg/well) or recombinant DIII of protein E of TBEV (27 μg/well). Nontreated cells were used as negative control,  for 6 hours at  37°C in the presence of 5% CO2. mRNA from human brain microvascular endothelial cells was isolated using RNeasy Mini Kit (Qiagen, Germany) according to manufacturer's instructions. 250 ng of RNA were reverse transcribed with oligodT primers using QuantSeq 3′ mRNA-Seq Library Prep Kit (Lexogen, Austria).
attributes.tag: Experimental Factor: stimulus
attributes.value: West Nile Virus protein E domain DIII
attributes.tag: Experimental Factor: time
attributes.value: 6