Fig. 8: Faf-mediated regulation of Hpo and Ft signalling is conserved.

a–d USP9X regulates Hpo signalling readouts in vivo. XY confocal micrographs of third instar wing imaginal discs carrying ex-lacZ (a, b) or DIAP1::GFP (c, d) and expressing GFP (a), lacZ^RNAi (c) or UAS-USP9X under the control of en-Gal4 (b,d), showing β-Gal antibody staining (a, b; red in a’, b’) or direct GFP fluorescence (c, d; green in c’, d’). Ventral is up in XY sections. GFP and RFP mark the hh-Gal4-expressing compartment (right) in a, b and c, d, respectively. Dashed lines depict anterior-posterior compartment boundary. Scale bar: 50 µm. e, f Quantification of Yki-mediated transcriptional reporter expression. Shown are posterior/anterior (P/A) ratios for ex-lacZ (e) and DIAP1::GFP levels (f). Data are shown as average ± standard deviation, with all data points represented. (n = 9, 12 in e, and n = 10, 13 in f). Significance was assessed using an unpaired two-tailed t-test, with Welch’s correction. ***, p < 0.001. g–o USP9X regulates tissue growth and modulates Ft-mediated growth phenotypes. Shown are adult wings from flies raised at 25°C expressing the indicated transgenes in the wing pouch under the control of nub-Gal4 (nub > ). Quantification of the effects of USP9X on growth is shown (i, l and o). Data are represented as % of the average wing area of controls (nub > GFP, average set to 100%). Data are shown as average ± standard deviation, with all data points depicted. (n = 22, 24 in I; n = 22, 21, 24 in l; and n = 22, 19, 23 in o). Significance was assessed using one-way ANOVA analysis comparing all genotypes to the nub > GFP control (black asterisks), with Dunnett’s multiple comparisons test. For pairwise comparisons, unpaired two-tailed t-tests with Welch’s correction were used. **, p < 0.01; ***, p < 0.001; ns, non-significant. Scale bar: 500 µm.