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In 2004 and 2005 normalising was performed annually.
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Background correction was applied using the "normexp" function and normalisation was performed using normalise between arrays "quantile" function.
Background subtraction was performed normalising the data to the negative control (bacterial plasmid pUC18).
The analysis was performed normalising against the calibrator and the result was the average of the three repeated experiments.
β-Galactosidase assay was performed to normalise for transfection efficiency.
An additional plate was set up concurrently and a crystal violet assay was performed to normalise ATP measurements to cell number.
To assess the effect of differentiation status and immune status on the expression profile, unsupervised hierarchical clustering of the GCRMA normalised and filtered probes (n=22 926) was performed using Ward average linkage.
Since the heart weight and coronary volume were subjected to intra- and inter-group variations, a correction was performed to normalise the input-function of the vasodilatation agents according to the coronary flow.
The relative transcript levels of the target genes were normalised against actin mRNA levels; quantification was performed using the comparative Ct method.
Data analysis was performed using the comparative Ct method normalised against β-actin expression.
Quantification was performed by the comparative Δ CT method normalising CT values to β-actin.
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