organism: Homo sapiens
description: Protocols: Human brain microvascular endothelial cells (hBMECs) were cultured in 6-well plates then incubated either with live N. meningitidis or recombinant MafA 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. N. meningitidis (NM, isolate M1/03, serogroup B, strain MC58).) was plated on commercially prepared Thayer Martin agar and a single isolated colony was transferred into 50 mL of Brain Heart Infusion Broth enriched with 10 mM MgCl2 . Bacteria were grown at 37°C and 5% CO2 until OD600= 0.6 (mid-exponential phase). Human brain endothelial cells cultured in 6-well plates were incubated either with live Neisseria meningitidis (MOI 0.5,bacterial cells were washed with minimal essential medium, Sigma) or recombinant MafA (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: ENA-FIRST-PUBLIC
attributes.value: 2019-06-15T04:02:31Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-05-30T17:04:53Z
attributes.tag: External Id
attributes.value: SAMEA5669586
attributes.tag: INSDC center name
attributes.value: Laboratory of Biomedical Microbiology and Immunology, The University of Veterinary medicine and Pharmacy in Kosice
attributes.tag: INSDC first public
attributes.value: 2019-06-15T04:02:31Z
attributes.tag: INSDC last update
attributes.value: 2019-05-30T17:04:53Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8008:Sample 6
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: hCMEC/D3
attributes.tag: cell type
attributes.value: blood vessel endothelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: organism part
attributes.value: brain
attributes.tag: sample name
attributes.value: E-MTAB-8008:Sample 6
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: stimulus
attributes.value: Neisserial ligand MafA
attributes.tag: time
attributes.value: 6