strain: C57BL/6J gender: male tissue: left ventricle of heart agent: control
Treatment protocol
After two weeks of acclimation, the mice were divided into the following five groups: (1) control, (2) 4.5 mg/kg/day pitavastatin calcium (Livalo tablet, Kowa, Nagoya, Japan), (3) 21.0 mg/kg/day pravastatin sodium (Mevalotin tablet, Daiichi-Sankyo, Tokyo, Japan), (4) 10.9 mg/kg/day rosuvastatin calcium (Crestor tablet, Shionogi, Osaka, Japan), and (5) 11.4 mg/kg/day atorvastatin calcium hydrate (Lipitor tablet, Astellas, Tokyo, Japan). Each statin tablet was powdered and given as a food admixture; the doses were calculated on the basis of daily food consumption (0.15 g/gBW). After four weeks, heart samples were obtained from each mouse under diethyl ether anesthesia.
Growth protocol
Male C57BL/6J mice (n = 20) were obtained from Charles River Laboratories Japan (Yokohama, Japan) at six weeks of age and maintained under specific pathogen-free conditions with controlled temperature and humidity and a 12-h light/12-h dark cycle. They were given standard chow (CE-2; CLEA Japan, Tokyo, Japan) and water ad libitum.
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from the left ventricle of each animal using an RNeasy Fibrous Tissue Mini Kit (Qiagen) according to the manufacturer's instructions.
Label
biotin
Label protocol
Amplified cDNA was prepared from 100 ng of total RNA using an Ovation RNA Amplification System V2 Kit (NuGEN Technologies) and purified using a PCR purification kit (Qiagen). According to the manufacturer’s instructions, 3.75 µg of purified cDNA was fragmented and labeled using an FL-Ovation cDNA Biotin Module V2 Kit (NuGEN Technologies).
Hybridization protocol
The fragmented and labeled amplified cDNA was hybridized to a GeneChip Mouse Genome 430 2.0 Array (Affymetrix) for 18 hr at 45C. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
Scan protocol
GeneChips were scanned using the GeneChip Scanner 3000 with Workstation (Affymetrix).
Description
n/a
Data processing
The CEL file data were analyzed with Microarray Suite version 5.0 (MAS 5.0) to generate CHP files (represented in Sample tables below). Subsequent data processing involved normalizing to the median expression value of all samples with GeneSpring GX version 11.0 software (Agilent Technologies).
