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These observations suggest that the missed responsive genes are more likely to connect with the biggest responsive module by transcriptional regulation rather than protein-protein interaction.
Among others, DEG in the F. graminearum responsive module B encoded glutathione S-transferases (GST), UGTs, glucanases, protein kinases and WRKY transcription factors (Additional file 11).
One of the more promising aspects of weighted gene coexpression network analysis was the identification of a common abiotic stress responsive module (red module) that enriched differentially expressed genes for all treatments investigated.
The discovery of this ankyrin family member as the hub in our common stress responsive module suggests that salicylic acid signaling may play a role in abiotic stress response, which would corroborate results from exogenously applied salicylic acid [ 52].
In addition, this common stress responsive module was enriched with genes known to participate in calcium and calmodulin signaling pathways, which have been shown to participate in a multitude of cellular functions including cell death [ 53].
This was done by ranking all modules from most to least responsive to CR (or aging) (i.e., rank modules by M), and discarding any module sharing at least one gene with any more responsive module of higher rank.
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The expressions of the Eigen genes were also tissue-specific because the responsive modules in root and shoot samples were mostly different for a selected condition.
In addition, specific stress responsive modules were identified.
The final step in the analysis was to screen all modules, and retain those modules most responsive to CR (or aging), while discarding less responsive modules that overlapped with more highly ranked modules.
The genes might be considered as hub genes to a specific stress with the most connected genes in the responsive modules developed from the WGCNA analysis [ 60, 61].
Furthermore, we demonstrate that more sophisticated regulatory motifs, such as small-molecule responsive modules can be built for crisprTFs, thus enabling external control of crisprTF-based transcriptional circuits.
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