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In brief, starting with the mouse datasets, we first partitioned the induced genes into disjoint groups according to the subset of stimulators to which the genes were responsive.
The first study, by Nau et al. [ 9], examined expression profiles in human monocyte-macrophages at several time points (1 hr, 2 hrs, 6 hrs, 12 hrs and 24 hrs) after stimulation by various agents; among them LPS and PIC are common to the stimuli examined by the mouse datasets we analyzed (this dataset is hereafter called HsM1).
Following the analysis applied to the mouse datasets, we partitioned the genes induced by LPS and PIC in the human datasets to the three kinetic sets: 'EE'ED'ED' and 'DD' (again, using the 2 hr time point as the boundary between early and delayed time points), according to the kinetics of their activation, and searched for over-represented signals in the promoters of these gene sets.
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Hence, for each human and mouse dataset we studied the expression of genes carrying bivalent domains in ESCs that were obtained from the overlap of three ChIP-on-chip studies in human [65], [66], [67] or in mouse [60], [65], [77] (Figure S2).
For the human-mouse dataset we observe that the number of true positives increases rapidly as α increases and then decreases at the same rate after reaching a maximum of 14133 when α is 0.53.
In our updated mouse KO dataset, we compiled 211 singleton mouse KO targeted genes with available protein connectivity data, as well as 845 mouse KO duplicates [ 26].
We began with the mouse datasets because they included more stimulators and denser kinetics than the human datasets.
In the complete mouse promoter dataset, we identified 29 significant motifs corresponding to matches to TRANSFAC matrices, 4 to JASPAR CORE matrices, 9 to JASPAR phyloFACTS, and 22 to 6mers.
In contrast to the results found on the mouse dataset (Table 3), we did not detect here an over-representation of ATF/CREB in the 'EE' set (representing early induction and peak).
In agreement with the results obtained on the mouse dataset, here too we observed a strong enrichment for NF-κB and ISRE elements in the 'ED' (early induction, delayed peak) and 'DD' (delayed induction and peak) sets, respectively (Table 5).
We combined 1,398 human and mouse datasets to computationally infer ISG modules and their regulators, validated by genetic analysis in both species.
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