Extended Data Fig. 4: Relationships between ATP levels, mitochondrial protonmotive force, and sleep in PNs and dFBNs.
a, Maximum-intensity projection of dFBNs expressing R23E10-GAL4-driven ATeam (top left, CFP channel). One dFBN has been filled with biocytin and a defined ATP concentration through a whole-cell electrode (bottom left). The right panel shows the least-squares fit of a Hill equation (dissociation constant 1.75 mM ATP, Hill coefficient 2.1) to the mean yellow-to-cyan emission ratio of biocytin-labelled dFBNs containing 0.15, 1.5, and 4 mM ATP (n = 3 cells each). b, Summed-intensity projections of PN dendrites expressing iATPSnFR plus RFP, in rested and sleep-deprived (SD) flies. Emission ratios are intensity-coded according to the key below and unaltered by sleep deprivation (P = 0.6616, two-sided t-test). c, Sleep in flies expressing R23E10-GAL4-driven Ucp4A or Ucp4C and parental controls (P ≤ 0.0139, Dunn’s test after Kruskal-Wallis ANOVA). d, A 400-ms pulse of green light elevates ATP in dFBNs expressing iATPSnFR plus tdTomato and mito-dR but not in dFBNs lacking mito-dR (n = 5 flies of either genotype, ∆p photogeneration effect: P < 0.0001, time × ∆p photogeneration interaction: P < 0.0001, two-way ANOVA). e, f, Sleep during the first 60 min after illumination (e, P ≤ 0.0279, Dunn’s test after Kruskal-Wallis ANOVA) and cumulative sleep percentages in flies expressing R23E10-GAL4-driven mito-dR, with or without retinal, and parental controls (f, ∆p photogeneration effect: P < 0.0001, time × ∆p photogeneration interaction: P < 0.0001, mixed-effects model). Asterisks, significant differences (P < 0.05) from both parental controls or in planned pairwise comparisons. Data are means ± s.e.m.; n, number of cells (a), antennal lobe glomeruli (b), or flies (c–f). Scale bars, 20 µm (a,b). For statistical details see Supplementary Table 2.
