normalization data transformation protocol
miRNA signal intensities from GeneView files were subjected to quantile normalization
array scanning protocol
Fluorescent signal intensities were detected by using the Agilent Scan Control 7.0 Software on an Agilent DNA Microarray Scanner at a resolution of 2 micron.
nucleic acid hybridization to array protocol
Slides were hybridized for 20 h at 55°C using an Agilent hybridization system.
nucleic acid labeling protocol
Total RNA samples (100 ng) containing miRNA were labeled with cyanine 3-pGp (Cy3) using the Agilent microRNA Complete Labeling and Hyb Kit.
Old mice were subdivided into two groups and fed different diets based on a modified AIN 76 diet: H-EVOO group was fed a diet in which the lipid component was provided by an extra-virgin olive oil high in phenolic antioxidants (718.8 mg/kg of total polyphenols); L-EVOO group by the same extra-virgin olive oil deprived of phenolic compounds (9.3 mg/kg of total polyphenols). Mice were sacrificed after 6 or 12 months of treatment. Male C57Bl/6J and TgCRND8 mice were sacrificed at 3-6 months of age. Brains were removed, dissected and immediately frozen in liquid nitrogen and stored at - 80 C.
Male C57Bl/6J mice aged 10 months at the beginning of the experiment were fed for 6 or 12 months with the experimental diets; young C57Bl/6J mice (4-6 months) and transgenic hemizygous TgCRND8 mice (3 months of age) were fed with a Standard lab chow.
nucleic acid extraction protocol
Total RNA was isolated from mice brain area by using the Absolutely RNA miRNA Kit (Agilent) according to the manufacturer’s protocol.