<i>Δ-cyc</i> expression in <i>Pdf</i>+ cells prevents sLN<sub>v</sub>s fasciculation.

<p>(A) Representative timeline of the experiments in the figure. Flies were kept in LD conditions at 28°C for their entire lifespan. Dissections were performed within days 6–8 post-eclosion at ZT2-3. (B) Representative brain confocal images of anti-PDF (green) and anti-RFP (magenta) staining in the sLN<sub>v</sub>s of flies in which <i>Δ-cyc</i> was constitutively expressed in the <i>Pdf</i>+ cells using a <i>Pdf-Gal4;tub-Gal80</i><sup><i>ts</i></sup> driver. Each line also included a <i>Pdf-RFP</i> transgene. White arrows indicate branching of sLN<sub>v</sub> dorsal projections (left), and dorsal termini of the sLN<sub>v</sub> projections (right) for the experimental genotype. The images are representative of two independent experiments. Scale bar = 50 μm. LN<sub>v</sub> morphology was quantified by comparing the sLN<sub>v</sub> projection length until the branching point (C), the total number of intersections of the sLN<sub>v</sub> ventral projections (D), the full sLN<sub>v</sub> projection length (E), and the total number of lLN<sub>v</sub> intersections (F). One-way ANOVA was used to analyze normally distributed data (C, F). For nonparametric data sets, a Kruskal-Wallis tests followed by Dunn’s multiple comparisons tests was used (D,E). * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars indicate SEM. Each dot corresponds to one brain. For each genotype: 9 ≤ n ≤ 12. (G-H) Behavioral phenotypes of constitutive Δ-<i>cyc</i> expression. Experiments were conducted at 28°C. (G) Population Activity (left) plots for flies during days 3–5 of the LD cycle at 28°C (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1011441#pgen.1011441.t001" target="_blank">Table 1</a> for additional quantifications). (H) Percent rhythmicity for the indicated genotypes under DD. R = Rhythmic and AR = arrhythmic. Fisher’s exact contingency tests were used to analyze the percentage of rhythmic flies under DD (DD1-8). *** p < 0.001. Error bars indicate SEM. For each genotype: 24 ≤ n ≤ 32.</p> <p>(PDF)</p>