|
| Status |
Public on Apr 03, 2018 |
| Title |
HMB-treated (24h) differentiated equine satellite cells_exposed to H2O2 (1h)_rep4 |
| Sample type |
RNA |
| |
|
| Source name |
HMB-treated (24h) differentiated equine satellite cells_exposed to H2O2 (1h)_rep4
|
| Organism |
Equus caballus |
| Characteristics |
Stage: 3rd day of differentiation
tissue: m. semitendinosus
|
| Treatment protocol |
After the 2nd day of differentiation equine satellite cells were pre-incubated with HMB for 24h (50 µM) and exposed to hydrogen peroxide (3mM, last 1h during 24h pre-incubation with HMB).
|
| Growth protocol |
Equine satellite cells were cultured in Primaria culture flask (BD, USA). The growth medium (10%FBS/10%HS/DMEM/AB) was changed every two days. On the 10th day of proliferation cells were transferred to Collagen I Cellware 6-well plate. After obtaining 80% of confluence proliferation medium was replaced by differentiation medium (2%HS/DMEM/AB). FBS - fetal bovine serum, HS - horse serum, DMEM - Dulbecco's Modified Eagle Medium, AB - antibiotics: 1% Penicillin-Streptomycin, 0.5% Amphotericin B.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated according to the protocol supplied with the miRNeasy Mini Kit (Qiagen, USA). RNA quantity was measured spectrophotometrically using NanoDrop (NanoDrop Technologies, USA). The analysis of final RNA quality and integrity was performed with BioAnalyzer 2100 (Agilent Technologies, USA). To ensure optimal data quality only RNA samples with RIN number ≥ 9,2 were included into the analysis.
|
| Label |
Cy3
|
| Label protocol |
MiRNA Complete Labeling and Hyb Kit (Version 2.3, December 2010) (Agilent) was used for labeling, according to manufacturer protocol.
|
| |
|
| Hybridization protocol |
For hybridization, Microarray Hybridization Chamber (Agilent, USA) and Hyb-Buffer (Agilent, USA) were used according to manufacturer protocol.
|
| Scan protocol |
Microarray Scanner (model G2565CA) with SureScan High-Resolution Technology (Agilent, USA). Microarray data were extracted, the background subtracted and LOWESS normalisation performed using the standard procedures contained in the Agilent Feature Extraction (FE) Software Version 10.7.3.1.
|
| Description |
Biological replicate 4 of 8. HMB-treated (24h) differentiated equine satellite cells_exposed to H2O2 (1h)_rep1
|
| Data processing |
The statistical analysis was performed using Gene Spring 13 software default protocol (Agilent Technologies, USA). The statistical significance of the differences was evaluated using student test (p<0.05) and fold change FC>1.3. A multiple testing correction was performed using Benjamini and Hochberg False Discovery Rate (FDR) ≤0.05.
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| |
|
| Submission date |
Oct 06, 2015 |
| Last update date |
Apr 03, 2018 |
| Contact name |
Tomasz Sadkowski |
| E-mail(s) |
tomasz_sadkowski@sggw.pl
|
| Phone |
+48225936263
|
| Fax |
+48228472452
|
| URL |
http://sadkowski.info
|
| Organization name |
Warsaw University of Life Sciences - SGGW
|
| Department |
Department of Physiological Sciences
|
| Street address |
Nowoursynowska 159
|
| City |
Warsaw |
| ZIP/Postal code |
02-776 |
| Country |
Poland |
| |
|
| Platform ID |
GPL20990 |
| Series (1) |
| GSE73779 |
MicroRNA expression data from β-hydroxy-β-methylbutyrate (HMB) pre-incubated differentiating equine satellite cells (ESC) exposed to hydrogen peroxide |
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