characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-05-16T17:03:07Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-16T15:05:08Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104699850
characteristics.characteristic: External Id
characteristics.text: Temple University Lewis Katz School of Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2018-05-16T17:03:07Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-16T15:05:08Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS2301233
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-6605:LPI 2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: endothelial cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000115
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: HAECs were collected using RNAeasy kit. Human aortic endothelial cells (HAECs) (Lonza, CC2535; Walkersville, MD) were cultured in medium M199 (Hyclone laboratories, Logan, UT) supplemented with 15% fetal bovine serum (FBS; HyClone); endothelial cell growth supplement (ECGS, 50μg /mL; BD Biosciences, San Jose, CA); heparin (50μg/mL); and 1% penicillin, streptomycin, and amphotericin (PSA; Invitrogen, Carlsbad, CA). HAECs were grown on 0.2% gelatin-coated flasks, plates, or dishes and experiments were performed at passage 9. 18-hour vehicle or lysophosphatidylinositol (16:0) treatment RNA was collected using RNAeasy kit. mRNA from HAECs was purified from total RNA using poly-T oligo-attached magnetic beads. The mRNA was first fragmented randomly by addition of fragmentation buffer. Then first strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Double-stranded cDNA was purified using AMPure XP beads (Beckman Coulter, Beverly, USA). Remaining overhangs of the purified double-stranded cDNA were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, NEBNext Adaptor with hairpin loop structure was ligated to prepare for hybridization. In order to select cDNA fragments of preferentially 150~200 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter). Finally, the final library was gotten by PCR amplification and purification of PCR products by AMPure XP beads. After library construction, diluting library to 1.5ng/ul with the preliminary quantitative result by Qubit2.0 and detecting the insert size by Agilent 2100. Q-PCR was used to accurately quantify the library effective concentration (> 2nM), in order to ensure the library quality.
characteristics.characteristic: description
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: aorta
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000947
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: LPI 2
characteristics.characteristic: title