Fig. 6. PYO-treated dTTC19 KO flies.

a ATP in 5-day-old male flies (dTTC19 KO) and controls (w¹¹¹⁸), after 12–24 h fasting (n = 3 × 10 individuals). Significant difference between KO and controls was at 24 h. Means ± SEM (two-tailed Student’s t test, **p < 0.01). b PYO toxicity assessed in WT flies (w¹¹¹⁸) after 1.0–200 pmol PYO injection into the hemolymph of male flies. The % of surviving flies was recorded at 24, 48, and 72 h. Kaplan–Meier (p < 0.0001), with Bonferroni-corrected Mantel–Cox log-rank was: 1 pmol vs. 100–200 pmol: p < 0.001; 10 pmol vs. 100–200 pmol: p < 0.001; 50 pmol vs. 100–200 pmol: p < 0.01, p < 0.001 respectively). c ATP levels in 5-day-old control (w¹¹¹⁸ and dTTC19 KO) and 1 pmol PYO (w¹¹¹⁸ PYO and dTTC19 KO PYO) flies after 24 h fasting (n = 3 × 10 individuals). PYO increased ATP production in dTTC19 KO flies to that of w¹¹¹⁸ control. Means ± SEM (two-tailed Student’s t test, **p < 0.01). d PYO partially recovered bang sensitivity in 5-day-old flies (dTTC19 KO PYO), vs. controls (w¹¹¹⁸; w¹¹¹⁸ injection control flies, and dTTC19 KO injection control flies). The % of flies reaching threshold distances (2.8, 5.6, 8.4, and 11.2 cm) is shown for the indicated genotypes, ±PYO injection. Means ± 95% CI (one-way ANOVA with post hoc on Tukey’s test: *p < 0.05; **p < 0.01; ***p < 0.001). e As in (d). Bang test in 12-day-old KO flies after 1 pmol PYO (dTTC19 KO PYO) vs. controls as above. The % of flies reaching the threshold distances are shown as above. Error bars mean 95% CI (one-way ANOVA with post hoc on Tukey’s test: *p < 0.05; **p < 0.01; ***p < 0.001). f Representative oxyblot and quantification of protein oxidation with 1 pmol PYO (w¹¹¹⁸ PYO and dTTC19 KO PYO) and relative controls as above (n = 2 independent experiments). g Pgc-1α expression assessed by qRT-PCR in w¹¹¹⁸ and dTTC19 KO flies untreated or treated with 1 pmol PYO (n = 3 biological replicates). Means ± SEM (one-way ANOVA with post hoc on Tukey’s test, *p < 0.05). h Toxicity associated with double injections (1 pmol PYO solution, n = 58 individuals vs. control solution, n = 53 individuals) assessed in WT male flies w¹¹¹⁸. The % of surviving flies was assessed at 24, 48, and 72 h after the second injection.