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Statistical support was computed using the GoStat web server (Beissbarth and Speed 2004), employing a stringent cutoff of P ≤ 0.001 and Benjamini correction for false positive detection.
Gene Ontology term enrichment in up and down regulated gene sets were assessed using the GOstat web tool [37].
Functional annotation of differentially expressed genes was performed using the DAVID web-server [ 69] and GOstat [ 70]..For GOStat analysis, all significantly down-regulated transcripts with at least one associated GO description (39), previously obtained with Blast2GO, were compared against a background of 4,692 GO-annotated DLPD entries, using the GOStat program with default settings.
The distance matrices were converted into heatmaps using the matrix2png web-server (Pavlidis et al. 2003).
Multiple sequence alignments (MSA) for the highly diverged AQPs were generated using the PROMALS3D web-server.
Nucleotide sequences were translated into protein sequences using the TRANSLATE web-server [ 65].
Homology searches against GenBank were done using the BLAST Web-server at NCBI [ 32].
The disulfide bond patterns were verified using the web-server BRIDGED [ 79, 80].
The enriched GO biological processes were clustered and visualized using the web-server REVIGO (http://revigo.irb.hr).
We calculated GO term enrichments using the GOstats package [46].
Analysis was carried out using the GoStats package, Bioconductor.
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