Fig. 1.
Requirement of ATX2 in LNvs for normal circadian behavior. (A) Effects of depleting ATX2 in pacemaker neurons on period length of circadian behavior. (Upper) Bar graph showing period length (Y axis), percentage of rhythmic flies and number of flies tested (in the bars). Error bars = SEM. TG4 is short for tim-GAL4, TD2 for tim-GAL4, UAS-dicer2, PD2 for Pdf-GAL4, UAS-dicer2 and PG80 for Pdf-GAL80. Dicer2 increases RNAi effects (15). (Lower) Double-plotted actograms of control and TG4/Atx2RNAi-1 flies showing the last day of the light-dark (LD) cycle and 5 days of DD. (B) (Upper) ATX2 is severely downregulated in circadian neurons expressing Atx2 dsRNA. Fly brains were dissected at Zeitgeber Time (ZT) 0 (ZT0 corresponds to the beginning of the light phase of the LD cycle) and immunostained with anti-ATX2, anti-PER and anti-PDF antibodies. (Lower) Quantification of ATX2 levels in sLNvs, lLNvs, LNds and DN1s. Between 11 to 20 neurons were quantified per data point. *** = P< 0.001 as determined by Tukey’s multiple comparison test after one-way ANOVA, n.s. = not significant at the 0.05 level, in both panel A and B.