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High confidence CHD1 and CHD2 peaks were calculated using model-based analysis for ChIP-seq (MACS) with a stringency cut off of 1 × 10-15 and binding sites within 200 bp or less were merged [ 37].
BLAST searches of the GSR dataset were based on a representative DNA binding domain from each of the ten major ERF subgroups (I-X) [ 58] using low stringency (cut off value = 10).
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Using the same stringency cut offs, only two genes, LCN2 and WRN2, were found on both lists.
The percentage overlap is above 60% even at the lower stringency cut-off of P≤10−4 (for p73), supporting the similarity between p63 and p73 binding, and a further indication that sites in this range are bona fide p73 targets.
These results indicate many p73 sites from the lower stringency cut-offs represent true p73 binding targets, consistent with the notion that transcription factor binding affinities in vivo represent a continuum, rather than simple presence or absence of binding [29], [30].
Here, the values for the low stringency cut-off were taken.
Of the 510 genes, 219 had significant female-biased expression (q < 0.5; a reduced stringency cut-off was used given microarray data tend to have higher variance).
To this end, an automated GO-annotation of the sequences of contigs and singletons that showed a significant similarity with a Uniprot entry was achieved using the Blast2go software [ 91], with a stringency cut-off of 1e-6.
Contigs with more than 5 questionable clones were split up and re-aligned with the DQ-er function in three steps of increasing stringency (cut-off values 1e-24, 1e-27, and 1e-30).
This was in concordance with RT-qPCR analysis that revealed a massive up-regulation in IL-10 mRNA (Fig. 6g).>> -wrap-foot> A minimum of 3 animals per group were independently analyzed (stringency cut-off: p < 0.0001 above controls) Functional parameters characterizing the cardiovascular system were measured 24 h, 14 days and 28 days after I/R with a pressure volume catheter (Fig. 8a h).
The high stringency cut-offs used in this figure highlight the most significantly shifted sites (Fig. 4B arrows).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com