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The ARHGAP19 ortholog RhoGAP54D is important for Drosophila melanogaster locomotion. (A) Schematic of the RhoGAP54D locus. Exons, white blocks; untranslated regions (UTRs), grey blocks; intron, line. Insertion site of the RhoGAP54D[CRIMIC-Gal4] element and exonic region targeted by RhoGAP54D shRNA are shown (red arrows). (B) Confocal images illustrating RhoGAP54D-driven membrane-tagged CD4:TdTomato expression in the adult male Drosophila brain and thoracic ganglion. Brain, yellow arrow points to projections close to the antennal mechanosensory motor center. Thoracic ganglion, red arrows point to isolated cell bodies. Note the CD4:TdTomato signal surrounding Bruchpilot-labeled (BRP-labeled) neuropil domains. (C) Patterns of locomotor activity in flies globally expressing RhoGAP54D shRNA (actin > RhoGAP54D shRNA) and driver/shRNA alone controls. White bar, lights on; black bar, lights off. (D and E) Number of beam breaks across 24 hours (D) or during ZT12-13 (E), a period of peak activity. n = 15–16. (F–H) Patterns of locomotor activity (F), total (G) and peak (ZT12-13; (H) beam breaks in adult flies harboring a RhoGAP54D GFP fusion allele and expressing deGradFP components, enabling degradation of the RhoGAP54D:GFP fusion protein, and a RhoGAP54D:GFP homozygote control. n = 18 and 24 respectively. (I–K) Patterns of locomotor activity (I), total (J) and peak (ZT12-13; (K) beam breaks in adult flies heterozygous or homozygous for the RhoGAP54D[KO] null allele. n = 16 and 20 respectively. Central line in dot plots, mean. Error bars: SEM. *P < 0.05, **P < 0.005, ***P < 0.0005, 1-way ANOVA with Dunnett’s post hoc test (D), Kruskal-Wallis test with Dunn’s post hoc test (E), Mann-Whitney U-test (G and K), or t test with Welch’s correction (H and J).
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