|
Status |
Public on Apr 29, 2014 |
Title |
Control_2h_rep2 |
Sample type |
RNA |
|
|
Source name |
hCMEC/D3
|
Organism |
Homo sapiens |
Characteristics |
tissue: Brain endothelium cell line: hCMEC/D3 treatment: control time: 2 h
|
Treatment protocol |
hCMEC/D3 cells were seeded onto collagen-coated tissue culture plate supplied by Greiner Bio-one (Gloucestershire, UK) and maintained for 72 h after confluence. Culture media was then changed to EGM-2 media without VEGF. Cells were left untreated or treated with TNFα and IFNγ at the concentrations and times indicated for each experiment.
|
Growth protocol |
hCMEC/D3 cells were cultured in EGM-2 MV medium (Lonza, Slough Wokingham, UK) and supplemented with the following components obtained from the manufacturer: 0.025 % (v/v) rhEGF, 0.025 % (v/v) VEGF, 0.025 % (v/v) IGF, 0.1 % (v/v) rhFGF, 0.1 % (v/v) gentamycin, 0.1 % (v/v) ascorbic acid, 0.04 % (v/v) hydrocortisone and 2.5 % (v/v) foetal bovine serum (FBS) (hereafter referred to as EGM-2 medium).
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA of hCMEC/D3 cells were isolated using miRNeasy mini kit (Qiagen, Sussex, UK) according to the manufacturer’s protocols. The quantity (NanoDrop 1000 spectrophotometer) and the quality (2100 Bioanalyzer, RNA 6000 Pico LabChip; Agilent, Palo Alto, CA, USA) of the total RNA were analyzed prior to determination of mRNA levels
|
Label |
Cy3
|
Label protocol |
Total RNA (100 ng) was labelled with pCp-Cy3 using T4 RNA ligase (GE healthcare, Amersham, UK)
|
|
|
Hybridization protocol |
Total RNA including miRNA (100 ng) was first dephosphorylated with calf intestine alkaline phosphatase (GE healthcare, Amersham, UK), denatured with dimethyl sulfoxide, and labeled with pCp-Cy3 to the 3’-ends using T4 RNA ligase (GE healthcare, Amersham, UK). The labeled RNAs were hybridized to Agilent human miRNA microarray (Santa Clara, CA). Design ID(AMADID)=021827. Standard Agilent’s miRNA microarray hybridization protocol
|
Scan protocol |
Standard Agilent scaning protocol
|
Description |
Gene expression contol 2h pair to T2F-T
|
Data processing |
standard Agilent Feature Extraction protocol
|
|
|
Submission date |
Feb 26, 2013 |
Last update date |
Aug 29, 2016 |
Contact name |
Miguel Alejandro Lopez-Ramirez |
E-mail(s) |
malopezramirez@ucsd.edu
|
Phone |
858-822-6487
|
Organization name |
University of California, San Diego
|
Department |
School of Medicine
|
Street address |
9500 Gilman Drive
|
City |
La Jolla |
State/province |
California/San Diego |
ZIP/Postal code |
92093 |
Country |
USA |
|
|
Platform ID |
GPL14767 |
Series (2) |
GSE44692 |
MiR-155 promotes blood-brain barrier dysfunction in neuroinflammation (part 1) |
GSE44694 |
MiR-155 promotes blood-brain barrier dysfunction in neuroinflammation |
|