|
| Status |
Public on Apr 27, 2017 |
| Title |
cardiac_tissue_Lina3 |
| Sample type |
RNA |
| |
|
| Source name |
cardiac_tissue_High_fat_diet_with_DPP4i _for_8weeks
|
| Organism |
Mus sp. |
| Characteristics |
tissue: cardiac tissue
gender: male
diet: high fat diet
agent: DPP-4 inhibitor
age: 12weeks
|
| Treatment protocol |
Mice were fed a high-fat diet or a normal chow for totally 8 weeks from 4 weeks age, and analyzed at 12 weeks of age. A DPP-4 inhibitor was administered within water at the concentration of 10 mg/kg/day for 8 weeks since 4 weeks of age.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the RNeasy Microarray Tissue Mini Kit (QIAGEN, Valencia, CA) following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.1 ug RNA using the One-Color Microarray-Based Gene Expression Analysis (Low Input Quick Amp Labeling) 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 |
600 ng of Cy3-labelled cRNA (specific activity >6.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 25 ul containing 10×Blocking buffer and 25×Fragmentation buffer following the manufacturers instructions. On completion of the fragmentation reaction, 25 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Mouse Genome Oligo Microarrays 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 (G2505C) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 3um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
gene expression of cardiac tissue from a high fat diet fed with DPP-4 inhibitor administration mice
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.1.1 (Agilent) using default parameters (protocol GE1-107_Sep09 and Grid: 074809_D_F_20150624) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Apr 26, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Masayoshi Suda |
| E-mail(s) |
Suda.Masayoshi@mayo.edu
|
| Organization name |
Niigata University
|
| Department |
Department of Cardiovascular Biology and Medicine
|
| Street address |
1-757 Asahimachi-dori, Chuo-ku
|
| City |
Niigata |
| ZIP/Postal code |
951-8510 |
| Country |
Japan |
| |
|
| Platform ID |
GPL21163 |
| Series (1) |
| GSE98226 |
Gene expression of cardiac tissues in dietary obese mice administered with a dipeptidyl peptidase-4 inhibitor |
|
