normalization data transformation protocol
Feature Extraction version 10.7 was used to filter, normalize, and calculate the signal intensities and Cy5/Cy3 ratios.
array scanning protocol
Axon Genepix 4000B microarray scanner at 5 micrometer resolution. Data from both channels were extracted using the Agilent Feature Extractor Software, Version 10.7
nucleic acid hybridization to array protocol
18 hour incubation at 65°C and 10 rpm in an Agilent Hybridization oven (Agilent p/n G2545A)
nucleic acid labeling protocol
We used the Agilent Two-Color Microarray-Based Gene Expression Analysis version 5.7 protocol
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.
The animals were randomly subdivided into two groups and fed different diets (about 3 grams/mouse per day).We used a modified AIN 76 diet, with lipids providing about 21% of the total energy, composed of (in grams/100 grams of diet): 44.3 sucrose, 10.0 olive oil, 21.0 casein, 14.0 corn starch, 5.4 cellulose, 3.7 AIN 76 Mineral mix, 1.1 AIN 76 Vitamin mix, 0.3 methionine, and 0.2 choline. 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 months of treatment. 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 months with the experimental diets. Young C57Bl/6J mice (4-6 months) were also used as reference.