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Fold change obtained by qRT-PCR correlated well with the direction of fold change obtained from the normalized intensity data, confirming the validity of the microarray gene expression patterns (Table S6).
Differential expression and direction of fold change when one or more probesets or probes exist on each microarray are summarized in Table S2 and complete details are presented in Datasets S2 and S3 (both described in Text S1).
With regard to the direction of fold changes of the verified genes, we found the least contradiction in TvN (217/1260 = 17.2%), less in Meta (117/321 = 36.5%), and the most, in Pre (42/82 = 51.2%).
We show that by using stringent data processing requirements differential expression as determined by RNA-Seq is concordant with both the Affymetrix and Illumina platforms in more instances than it is concordant with only a single platform, and that instances of discordance with respect to direction of fold change were rare.
All qPCR results were concordant compared to microarray in terms of the direction of fold changes.
The direction of fold change was also determined for each gene at each time point comparison.
Similar(43)
Co-directed expression, i.e., identical direction of fold-changes in the two organisms, can be used as an important data filtering criterion: co-directed over expression only, co-directed repression only, both together, both anti-directed directions (under-over and over-under expressions).
Fourteen genes demonstrated consistent changes in gene expression (concordant direction of fold-change) by qRT-PCR when comparing mild and moderate COPD samples.
The direction of fold-change was consistent with the microarray data for each of the nine genes selected for RT-qPCR validation.
For deltamethrin, the direction of fold-change for c-fos and Egr1 is down in most dose groups measured by qRT-PCR.
Enrichment was scored assuming either non-directionality (disregarding direction of fold-change) or directionality (up- and down-regulated genes cancel out) and the best scoring method was retained for each functional category.
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