|
| Status |
Public on Apr 29, 2014 |
| Title |
Control_24h_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
hCMEC/D3
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Brain endothelium
cell line: hCMEC/D3
treatment: control
time: 24 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 24h pair to T24E-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 |
|
