Stat92E functions in glia to promote axon clearance and regulate Draper expression.

<p>(A) Z-stack confocal images of OR85e⁺ axons labeled with GFP in control (<i>OR85e-GFP/+; repo-gal4/+</i>) or <i>stat92e^RNAi</i> (<i>stat92e^RNAi</i>/<i>OR85e-GFP;repo-Gal4</i>/+) backgrounds. Uninjured and 5 days after maxillary palp ablation are shown. Arrows indicate position of maxillary nerves. (B) Quantification of GFP intensities in 85e⁺ glomeruli from (A), <i>p</i>-values were calculated using Student's <i>t</i> test. Error bars represent SEM. (C) Quantification of maxillary nerves with GFP⁺ axon fibers from (A). (D) Single slice confocal images; Draper antibody stains in adult brains of control (<i>repo-gal4</i>/+) and <i>stat92e</i>^RNAi (<i>stat92e</i>^RNAi/+;<i>repo-gal4</i>/+) backgrounds. Uninjured, one day after maxillary palp injury and one day after antennal ablation are shown. Dotted circles and boxes represent regions used for quantification of Draper immunoreactivity. (E) Quantification of data from (D), <i>p</i>-values were calculated using one-way ANOVA followed by Tukey's post hoc test, <i>n.s.</i>, not significant. Error bars represent SEM. (F) Western blot analysis of Draper protein levels in ∼3 adult central brain regions of control (<i>repo-gal4</i>/+), <i>stat92e^RNAi</i> (<i>UAS-stat92e^RNAi/+; repo-gal4/+</i>), and <i>draper^RNAi</i> (<i>draper^RNAi</i>/+;<i>repo-gal4/+</i>) genotypes. α-Tubulin was used as a loading control. Full blot located in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001985#pbio.1001985.s003" target="_blank">Figure S3</a>. (G) Quantification of Draper protein levels shown on Western blot after normalization to α-Tubulin, <i>n</i> = 3, <i>p</i>-values were calculated using one-way ANOVA followed by Tukey's post hoc test. (H) Real-time PCR analysis of <i>draper</i> mRNA levels in adult central brain region of control (<i>repo-gal4/+), drpr^RNAi</i> (<i>draper^RNAi</i>/+;<i>repo-gal4/+), pi3k92e^CAAX(UAS-pi3k92e^CAAX/+;repo-gal4/+), pi3k92e^RNAi(UAS-pi3k92e^RNAi/+;repo-gal4/+), and stat92e^RNAi</i> (<i>stat92e^RNAi</i>/+;<i>repo-gal4/+) genotypes</i>. Draper Ct values were normalized to ribosomal protein L32 and results are presented as fold induction relative to control. Values represent mean ± SEM from three independent RNA isolations. <i>p</i>-Values were calculated using one-way ANOVA and Dunnett's post hoc test.