|
| Status |
Public on Oct 01, 2018 |
| Title |
INS1_cells_6h_Giv_Ctrl_replicate_2 |
| Sample type |
RNA |
| |
|
| Source name |
INS1 cells, treated with Givinostat, 6h
|
| Organism |
Rattus norvegicus |
| Characteristics |
cell-line: Rat pancreatic INS1 cells
group: 6h_Giv_Ctrl
|
| Treatment protocol |
INS1 cells were incubated for 48h prior to exposure with or without 125 nM Givinostat (ITF2357) for 1 h, prior to addition of 150 pg/ml IL-1-β and 0.1 ng/ml IFN-γ for 6-24 h
|
| Extracted molecule |
total RNA |
| Extraction protocol |
MiR-enriched total RNA was purified with the miRCURY™ RNA Isolation Cell and Plant Kit (Exiqon)
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent SurePrint 8x15k Rat miRNA Agilent microarrays (G4473C) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B)
|
| Description |
miRNA expression data from INS1 cells in 6h_Giv_Ctrl group
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Jul 20, 2018 |
| Last update date |
Oct 01, 2018 |
| Contact name |
Flemming Pociot |
| E-mail(s) |
flemming.pociot@regionh.dk
|
| Organization name |
Steno Diabetes Center Copenhagen
|
| Department |
T1D Biology
|
| Street address |
Niels Steensens Vej 2
|
| City |
Gentofte |
| ZIP/Postal code |
2820 |
| Country |
Denmark |
| |
|
| Platform ID |
GPL22760 |
| Series (1) |
| GSE117451 |
MicroRNAs and histone deacetylase inhibition-mediated protection against inflammatory β-cell damage |
|
