|
| Status |
Public on Dec 31, 2022 |
| Title |
Liver_FGF21_low_03 |
| Sample type |
RNA |
| |
|
| Source name |
Liver FGF21 low group
|
| Organism |
Bos taurus |
| Characteristics |
breed: Holstein
gender: female
condition: FGF21 low
tissue: liver
|
| Treatment protocol |
All animals received the same TMR, which was sufficient to meet requirements of the cows during dry period and lactation, respectively.
|
| Growth protocol |
The experiment with multiparous Holstein cows was carried out at the Educational and Research Centre for Animal Husbandry Hofgut Neumühle (Münchweiler an der Alsenz, Rhineland-Palatinate, Germany). All procedures described in this study were performed according to the German Animal Welfare Act. The animal experiment was approved by the local Animal Care and Use Committee (Provincial Government of Coblenz, Germany, 23 177–07/ G15–20–040725_M).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from frozen liver aliquots (15-20 mg) was isolated using TRIzol reagent (Invitrogen, Karlsruhe, Germany) according to the manufacturer’s protocol. RNA quality was checked with an Agilent 2100 Bioanalyzer using the RNA 6000 Nano Kit (Agilent Technologies Inc., CA, USA). Samples were processed at an Affymetrix Service Provider and Core Facility, “KFB-Center of Excellence for Fluorescent Bioanalytics” (Regensburg, Germany; www.kfb-regensburg.de) as previously described. The RNA integrity number (RIN) value for all liver biopsy samples was 6.02 ± 0.54 (mean ± SD).
|
| Label |
Biotin
|
| Label protocol |
200 ng of total RNA was used to generate double-stranded cDNA. 12 µg of subsequently synthesized cRNA were purified and reverse transcribed into single-stranded (ss) cDNA, whereby unnatural dUTP residues were incorporated. Purified ss cDNA was fragmented using a combination of uracil DNA glycosylase (UDG) and apurinic/apyrimidinic endonuclease 1 (APE 1) followed by a terminal labeling with biotin.
|
| |
|
| Hybridization protocol |
3.8 µg fragmented and labeled ss cDNA were hybridized to Affymetrix Bovine Gene 1.0 arrays for 16 h at 45° C and 60 rpm in a GeneChip hybridization oven 640.
|
| Scan protocol |
Hybridized arrays were washed and stained in an Affymetrix Fluidics Station FS450, and the fluorescent signals were measured with an Affymetrix GeneChip Scanner 3000 7G. Fluidics and scan functions were controlled by the Affymetrix GeneChip Command Console v4.3.3 software.
|
| Description |
P232_11_1201
|
| Data processing |
Summarized probe set signals in log2 scale were calculated by using the GCCN-SST-RMA algorithm with the Applied Biosystems GeneChip Expression Console v1.4 Software. After exporting into Microsoft Excel, average signal values, comparison fold changes (FC) and significance P-values were calculated.
|
| |
|
| Submission date |
Nov 28, 2022 |
| Last update date |
Dec 31, 2022 |
| Contact name |
Denise Gessner |
| E-mail(s) |
denise.gessner@ernaehrung.uni-giessen.de
|
| Organization name |
JLU Giessen
|
| Department |
Animal Physiology and animal nutrition
|
| Street address |
Heinrich-Buff-Ring
|
| City |
Giessen |
| ZIP/Postal code |
35392 |
| Country |
Germany |
| |
|
| Platform ID |
GPL16500 |
| Series (1) |
| GSE218916 |
Effect of high and low FGF21 expression on metabolic, inflammatory and antioxidative stress-related parameters in early lactating dairy cows |
|
