nejire regulates PDFR respecification in l-LNvs of both young and mature flies.

(A) The amplitude of l-LNv responses to PDF on day 0 in Pdf-GAL4>UAS-Epac1 flies crossed to UAS-RNAi lines of the depicted transcription factors (ANOVA; F_[8,127] = 11.1, p = 1.26^E-11, *p < 0.05, modified Bonferroni test, n are listed below the x-axis). (B) PDF amplitude in control and 5-day-old sleep restricted Pdf-GAL4>UAS-Epac1 flies compared to Pdf-GAL4>UAS-Epac1 flies expressing UAS-nej^RNAi (n = 6–19 neurons/genotype). (C) In the absence of sleep loss, the l-LNvs of Pdf-GAL4>UAS-Epac1/UAS-nej^WT respond to PDF while age-matched Pdf-GAL4>UAS-Epac1 do not (t test, DF 34–1, t = 22.6, p = 3.74^e-5, n = 9–26 neurons/genotype). (D) Data quantified from (B). (t test, DF 40–1, t = 17.8, p = 4.10^e-4, n = 16–26). (E) The amplitude of l-LNv responses to PDF in l-LNvs on day 0 in Pdf-GAL4>UAS-Epac1 flies crossed to UAS-RNAi lines of the depicted cell surface receptors (ANOVA F_[10,105] = 5.38, p = 2.8⁻⁶ p < 0.05, modified Bonferroni test, n are listed below the x-axis). (F) The amplitude of l-LNv responses to PDF in l-LNvs on day 5 in sleep-restricted Pdf-GAL4>UAS-Epac1 flies crossed to UAS-RNAi lines of the depicted cell surface receptors (ANOVA F_[10,179] = 7.04, p = 3.3^E-9 p < 0.05, modified Bonferroni test, n are listed below the x-axis). Data underlying this figure can be found in S6 Data. FRET, Förster Resonance Energy Transfer; l-LNv, large ventral lateral neuron; PDF, pigment dispersing factor; PDFR, pigment dispersing factor receptor.