Nub-PB and Nub-PD regulate the same genes in an antagonistic and Relish-dependent manner.

<p>(A) Quantification of relative mRNA expression from selected AMP genes along with each isoform of <i>nub</i> (inserts), from whole fly extracts using RT-qPCR following overexpression of <i>nub-RB</i> (yellow bars) or <i>nub-RD</i> (magenta bars) using <i>c564>Gal4</i> (means ± SE, <i>N</i> = 3). (B) Quantification of relative mRNA expression from selected AMP genes along with each isoform of <i>nub</i> (inserts), from whole fly extracts using RT-qPCR following overexpression of <i>nub-RB</i> (yellow bars) or both (purple bars) using <i>c564>Gal4</i>^<i>ts</i> (means ± SE, <i>N</i> = 4). (C) Transfection of mbn-2 cells with a pA10-<i>CecA1-luc</i> construct (15) with or without expression plasmids for <i>nub-RB</i> and/or <i>Rel</i>. Cells were subsequently stimulated with bacterial peptidoglycan (in the form of crude LPS) to induce a robust immune response. The graph depicts mean values of relative luciferase activity ± SE (<i>N</i> = 3). (D-F) Quantification of relative mRNA expression from selected AMP genes using RT-qPCR following c564-driven overexpression of <i>nub-RB</i>, with or without a Rel mutant background (<i>Rel</i>^<i>E20</i>, orange bars) compared to controls and with or without systemic infection with <i>E</i>. <i>cloacae</i> (means ± SE; <i>N</i> = 3). Lower panels display proximal promoter regions of <i>DptA</i>, <i>CecA1</i> and <i>Drsl2</i>. Oct (ATSBAAAW; ●), and Oct-like (ATTCAAAT; ) motifs are depicted in the proximal promoter region of each gene [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006936#ppat.1006936.ref015" target="_blank">15</a>]. κ = κB-site; S = Stat92E-site; A = AP-1 site. Bars represent means ± SE. Asterisks and distinct letters denote significant differences (*<i>p<0</i>.<i>05</i>; **<i>p<0</i>.<i>01</i>; ***<i>p<0</i>.<i>001</i>; distinct lettering, <i>p</i><0.05).