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We measured fold changes in expression in two ways: within strains and between strains.
Between strains, we measured fold changes in gene expression within treatments and timepoints (as described in the following).
However, as expected there was not any linear correlation between the EDMA fold changes for the selected DMRs and their corresponding measured fold changes after the pyrosequencing confirmation (see Additional file 1: Figure S3).
The statistical robustness of the measured fold changes in the arrest data set was determined using a one-way ANOVA and a p-value cutoff of 0.05 was implemented.
Measured fold changes for three unique phosphorylated α-casein peptides (one singly phosphorylated peptides was identified in two different charge states) varied from 2.89 to 3.57, which an average fold-change of 3.11 and an RSD of 10.2%.
Within strains, we measured fold changes in gene expression after heat treatments relative to control treatment (e.g. PT relative to C, PT+HS relative to C, see Figures 2, 3 and 4).
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In addition to a high concordance with in vitro measured fold change, which was the primary aim of model design, the models per drug show good predictivity of therapy response for regimens including that drug, even in the absence of other clinically relevant factors such as background regimen.
For example, if the RMS deviation is 0.15 (15%), then a measured fold change between samples of 1.15 is a difference of one standard deviation.
Measured fold change in gene expression between two samples was used to set the color of the nodes, green indicating up regulation, red indicating down regulation and yellow indicating no change between samples.
Bantscheff et al. (2008) have thoroughly investigated this problem for iTRAQ labeling and concluded that the measured fold change is increasingly deviating from the expected ratio at broader isolation widths, thus indicating that the presence of co-eluting peptides significantly affects the reporter intensities.
Real-time quantitative PCR (qPCR) can be quite sensitive, but its reliance on relative differences in crossing thresholds rather than direct quantification makes it more suitable for measuring fold changes rather than absolute deletion frequencies (He et al., 2002; Chabi et al., 2003).
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