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Four independent pooled sets of samples were used for microarray (n = 4 biological replicates) analysis.
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Samples used for microarray analysis (n = 16) were taken from males (n = 5) aged 3-7 months.
Gene-specific real-time PCR validation of microarray was performed in an identical manner for H3K9 acetylation and DNA methylation enrichment[21] for the same subjects used for microarray experiments (n = 3/group; see Methods S1 for details).
Real-time PCR was performed using the same RNA samples used for microarray hybridization (n = 16).
Real-time PCR was performed using all RNA samples used for microarray analysis (N = 29).
RNAs determined to be of high purity (260/280 ~1.8) and intact by gel analysis were used for microarray analysis (n = 28) or quantitative real time RT PCR.
This discrepancy may be due to the small sample size (n = 43) used for microarray assay [ 16].
These analyses used the same RNA samples as those used for microarray profiling plus two additional biological replicates (n = 6).
Twelve samples from hyperoxia-selected adult flies (SO2A in 90% O2 chamber, n = 6) and control flies (in room air chamber, n = 6) were used for microarray experiments, in which had a pool of 25 male and 25 female flies from each chamber.
Total RNA isolated from adipose tissue, liver, muscle, and pancreas of a subset of F2 mice (n = 16) were used for microarray analysis, requiring 64 arrays.
The study includes samples used for microarray hybridization and samples from independent treatments; a total of n = 8.
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