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We confirmed that CecA1 and Dipt were also up-regulated in nub 1 mutants after treatment with antibiotics, excluding that excessive expression in nub 1 mutants is caused by ongoing infections.
Expression of CecA1 and Diptericin (Dipt) was significantly higher in nub 1 than in wt flies, whereas Drosomycin (Drs) expression was only slightly increased.
A few Toll pathway components were found to be down-regulated in nub 1 mutants, such as Persephone, GNBP3 and WntD.
From this global mRNA analysis, we conclude that numerous genes involved in immune defense reactions are abnormally expressed in nub 1 mutant flies.
The whole genome analysis in nub 1 mutants revealed a comprehensive picture of the role of Nub-PD in regulation of genes involved in the fly's immune system.
We first analyzed expression of four AMP genes, CecA1, CecC, Dipt and AttC, in nub 1 ; UAS-nub control flies, confirming strong over-expression of all four genes.
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These cells also express puckered, a reporter of JNK activity, in nub-Gal4 UAS-hppy /puc-lacZ discs (Fig. 4E F).
We confirmed that DfH99, DIAP and Puc over-expression efficiently suppressed the expression of activated Cas3 in nub-Gal4 UAS-CG7097/+ UAS-CG7097/+Fig. 4C–D and data not showing
The reduction in the number of cells in wings over-expressing hppy varies from 15% to 28% in nub-Gal4 UAS-hppy /+ heterozygous compared to nub-Gal4 UAS-CG7097 homozygous wings (Fig. 3H and Table S1).
While the two enhancers both contain the conserved DNA sequence TGCTGCTGTTG, the 11-mer is present twice in nub-31 and once in nub-32a.
In particular, the putative Hb/Cas docking site was detected multiple times within conserved sequences in nub-8 (2 sites), nub-9 (6 sites), nub-10 (2 sites), and nub-12 (5 sites) (Fig. 11b, c).
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