Fig. 5

foxq1a^bcz11 and foxq1b^bcz18 mutants exhibit wildtype transcriptional response to E. coli challenge in the larval brain.

a Graphical representation of the brain challenge assay. E. coli was injected into right brain tectum, and at 6 hours post injection (hpi), injected larvae were individually processed for RNA extraction, genotyping, cDNA synthesis, and qPCR analysis. b Bar chart shows significant upregulation of immune activation genes il1β, irg1, mpx, and tnfα after bacteria challenge at 6 hpi in wildtype larvae compared with uninjected controls. Water injected wildtype larvae exhibit no induction of activation genes, whereas LPS injected wildtype animals have a significant upregulation of il1β and irg1. After microbial activation by E. coli or LPS, a significant downregulation of foxq1a was found by about a factor of 2. No significant change was found for foxq1b after immune challenge. Data from wildtype injections validate the efficacy of the assay to activate the innate immune system. Dotted line marks no fold difference at 1. At least 6 or more independent biological samples were measured per category. c Bar plot shows transcriptional changes of target genes after E. coli injection in the brain at 6 hpi for control siblings and foxq1a and foxq1b mutants. n = 9–14 independent biological samples were measured per genotype. All error bars show standard error of means. Statistical significance in b and c was determined by multiple unpaired t-tests comparing between uninjected or sibling controls and the experimental groups with correction for multiple comparisons using the Sidak-Bonferroni method and without assuming equal variance. n.s., no statistical significance as defined by p > 0.05. *, p < 0.05; **, p < 0.01; ***, p < 0.001.