Supplementary Figure 4: Riquiqui and Minibrain affect wing growth independently of Dachs.

Wing imaginal discs from third instar D. melanogaster larvae containing clones of UAS-riqi1 (a) or UAS-mnbi1 (b) were stained for Dachs (grey in a and b). Clones were positively marked by co-expression of GFP (a’ and b’). Merged images are shown in (a” and b”). X-Z sections through the wing pouch are shown below planar sections of wing discs in a–b”. Dashed lines represent the section used to obtain X-Z images. No changes in Dachs expression or localization were observed. (d–e”) All images were recorded in pupal scutellum tissue (right-sided hemi-scutellum) at 20 hour after pupa formation. Anterior is to the right, the midline is to the top. Yellow circles mark the macrocheatae. Spatial information on the Fj-Ds gradient is shown in c as in ref. 1. Flip out clones of UAS-riqi and UAS-mnbi were generated and marked in red (d” and e” respectively). White boxes are magnified in d’ and e’. Yellow dots mark the mutant cells abutting wt cells (d’ and e’). In these Flip-Out clones no clear reduction of D:GFP levels was observed when compared with the surrounding tissue. There is also no clear repolarization of D:GFP near clone boundaries. (f) Wing area from adult female flies expressing UAS-EYFP as control, UAS-riqi, UAS-dachs RNAi (di), UAS-riqi concomitantly with UAS-di, under the control of rn-Gal4 (n = 20 for each genotype). Representative images of each genotype are shown. Data is presented as mean±SEM, ^*** = p<0.001. These results suggest Dachs and Riq control tissue growth by acting in parallel. Scale bars represents 50 μm in (a) and (b), 10 μm in (c), (d) and (e), and 200 μm in (f).