title: Sample 6
organism: Homo sapiens
description: Protocols: Human umbilical vein endothelial cells (HUVEC), purchased from Lonza (Cologne, Germany), were cultured in endothelial basal growth media (EBM) from Lonza, supplemented with hydrocortisone (1 µg/ml), bovine brain extract (12 µg/ml), gentamicin (50 µg/ml) human epidermal growth factor (10 ng/ml) and 10 % fetal calf serum. To simulate an unhealthy diet cells were seeded into cell culture dishes and incubated with the cultivation media for two days. Subsequently cells were treated with media, which was supplemented with either 0.4 mg/ml fructose (con) or 0.4 mg/ml fructose and 1 mg/ml LDL (high LDL). This media was refreshed every second to third day up for 5 to 7 days depending on the experiment. Total RNA-Isolation was performed by using the RNeasy kit from QIAGEN (Hilden, Germany). Cells were lysed using trizol and total RNA was precipitated by adding isopropanol. After centrifugation of this mixture, the liquid phase (RNA) was transferred to a QIAspin column. Subsequently RNA was washed and eluted following the manufacture guidelines. The RNA concentration was measured using a NanoDrop 2000c from Thermo Fisher (Schwerte, Germany) and RNA integrity as well as purity was analyzed by agarose gel electrophoresis. 1. Normalization: Dilution to 200 ng in 50 μL 2. Library Preparation: 50 μL of total RNA sample input. The Library preparation was performed according to the TruSeq® Stranded mRNA Sample Preparation Guide (Part # 15031047 Rev. E) on Sciclone NGS roboter (PerkinElmer).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-02-26T17:02:06Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-02-01T13:36:43Z
attributes.tag: External Id
attributes.value: SAMEA5315027
attributes.tag: INSDC center name
attributes.value: Heinrich-Heine University Duesseldorf
attributes.tag: INSDC first public
attributes.value: 2019-02-26T17:02:06Z
attributes.tag: INSDC last update
attributes.value: 2019-02-01T13:36:43Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7647:Sample 6
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: HUVEC
attributes.tag: cell type
attributes.value: endothelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: growth condition
attributes.value: high LDL
attributes.tag: organism part
attributes.value: umbilical vein
attributes.tag: sample name
attributes.value: E-MTAB-7647:Sample 6
attributes.tag: scientific_name
attributes.value: Homo sapiens