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. 2016 Jan 29;7:10509. doi: 10.1038/ncomms10509

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(a) Myoblast area and number are not altered. Confocal images of late third instar wing discs stained with anti-β-GAL (twi-lacZ), anti-Mef2 antibody and DAPI. Z-stack-projected images of adepithelial area were magnified. Quantification of total number of nuclei in Twi-positive area using DAPI staining, and Twi-positive area relative to total wing area, mean±s.e.m., n=14, 10 wing discs per genotype, t-test with Welch's correction, P>0.01. Two independent experiments. (b) The expression of myoblast-specific genes in wing discs is not affected. Expression of twi, Holes-in-muscles (Him) and Mef2 genes quantified by RT–qPCR. Arp53D was de-repressed in Dp-depleted wing discs. Mean±s.e.m., N=3 and 5 samples per genotype, two-way analysis of variance (ANOVA), P>0.05 (left panel) and Mann–Whitney test, *P<0.05 (right panel). (c) The onset of myotube differentiation proceeds normally. Developing DLMs (asterisks) at 21 h APF stained with the reporter vg^AME-lacZ to mark the onset of myotube formation and anti-Mef2 antibody. Arrowhead points to a fusion event. Anterior is to the top. Quantification of the total number of Mef2-positive nuclei per image (μm²), the area of Mef2-positive nuclei (μm²), mean±s.e.m., N=9 thoraces per genotype, t-test with Welch's correction, P>0.05. Three independent experiments. (d) The expression of myogenic genes in developing flight muscles at 21 h APF is normal. The genes measured are blown fuse (blow), singles bar (sing), myoblast city (mbc), Holes-in-muscles (Him), twist (twi), lame duck (lmd), erect wing (ewg), vestigial (vg), Mef2, Act88F, htl and stumps. mean±s.e.m., N=4 independent samples per genotype, two-way ANOVA, P>0.05. (e) Myoblast fusion to developing DLMs is not grossly altered. Developing DLMs at 40 h APF stained with Phalloidin to mark early myofibrils and anti-Mef2 antibody. Anterior is to the top. Quantification of the total number of Mef2-positive nuclei per area (μm²), the area of Mef2-positive nuclei (μm²), Mean±s.e.m., N=8 thoraces per genotype, t-test with Welch's correction, P>0.01. Two independent experiments. Genotypes are Mef2-GAL4/UAS-mCherry-RNAi and UAS-Dp-RNAi;Mef2-GAL4. Scale bar, 50 μm (a–c), and 20 μm (e).