Extended Data Fig. 9: Inducing mitochondrial fission or fusion in dFBNs alters arousal thresholds without causing overexpression artefacts or anatomical defects.
a, b, Percentages of flies awakened by mechanical stimuli lasting 0.5 s (left panels), 2 s, or 20 s (right panels). The average percentages awakened by 0.5-s stimuli in the left panels are reproduced on the right. Manipulations that increase fission fail to lower the arousal threshold, possibly because of a floor effect linked to the R23E10-GAL4 strain (a, P ≥ 0.3354 relative to ≥1 parental control, Dunn’s test after Kruskal-Wallis ANOVA). With the exception of the overexpression of Marf alone (P > 0.9999), manipulations that increase fusion raise the arousal threshold (b, P ≤ 0.0371, Dunn’s test after Kruskal-Wallis ANOVA). c, Sleep in flies expressing R23E10-GAL4-driven Marf is insensitive to the co-expression of fluorescent proteins in the cytoplasm (tdTomato) or the outer mitochondrial membrane (OMM-mCherry) (P ≥ 0.1106, Dunn’s test after Kruskal-Wallis ANOVA), in contrast to the synergistic effect of overexpressing Opa1 (Fig. 4c, Extended Data Fig. 8a). d, Maximum-intensity projections of dFBNs in flies carrying R23E10-GAL4-driven overexpression constructs or RNAi transgenes targeting the mitochondrial fission or fusion machinery. Five brains per genotype were imaged; representative examples are shown. Data are means ± s.e.m.; n, number of flies; asterisks, significant differences (P < 0.05) from both parental controls. Scale bar, 100 µm (d). For statistical details see Supplementary Table 2.
