Fig. 7. WAKE modulates male-male courtship behaviour via IIS-regulated JH biosynthesis in the CA.

a Over-expression of InR^CA in the CA effectively evoked male-male courtship behaviour. WAKE downregulation in IPCs simultaneous expression of dominant negative-InR or chico dsRNA in the CA significantly suppressed male-male courtship behaviour. n = 18 for each test group no. 1–6, n = 24 for each test group no. 12–16. p < 0.0001, Kruskal–Wallis test. ^***p < 0.005 and ^****p < 0.0001, post hoc Dunn’s multiple comparisons test. n = 18 for each test group no. 7–11, p < 0.0001, one-way ANOVA using F-test, post hoc Tukey’s multiple comparisons test, ^***p < 0.005 and ^****p < 0.0001. b After eclosion, wake³²⁰⁹⁹ treated with precocene I significantly reduced male-male courtship behaviour. n = 15 for test group no. 1 and 2. ^****p < 0.0001, two-tailed Mann–Whitney U-test. WAKE-deficient in IPCs and simultaneous expression of jhamt or hmgcr dsRNA in the CA significantly suppressed male-male courtship behaviour. n = 18 for test group no. 3–7, p < 0.0001, one-way ANOVA using F-test, post hoc Tukey’s multiple comparisons test, ^**p < 0.01, ^***p < 0.005 and ^****p < 0.0001. n = 24 for test group no. 8–12. p < 0.0001, Kruskal–Wallis test. ^****p < 0.0001, post hoc Dunn’s multiple comparisons test. wake³²⁰⁹⁹ males treated with precocene I or simultaneous expression jhamt or hmgcr dsRNA in the CA did not affect male-female courtship behaviour. n = 19, 18 for test group no. 13 and 14. p > 0.05 (n.s.), two-tailed Mann–Whitney U-test. n = 18, 18, 17 (from left to right) for test group no. 15–17. p = 0.0413, Kruskal–Wallis test, p > 0.05 (n.s.), post hoc Dunn’s multiple comparisons test. c Relative expression of the juvenile responsive-gene kr-h1 via qPCR, n = 3 for each. Scatterplots include ± SEM. Source data are provided as a Source Data file.