Fig. 5

Knockdown of candidate genes is affected by genetically altered DmManf level. Overexpression of DmManf (a) and heterozygous DmManf ^Δ96 mutant background (b) affect phenotypes of candidate gene knockdown with wing driver MS1096-GAL4 (a and b in a, a in b) and semi-ubiquitous driver 69B-GAL4 (c and d in a, b in b). The knockdown of COQ7 and CG9249 are discussed in the main text. Knockdown of Spn27A (Serpin-27A, an inhibitor of serine-type endopeptidase activity) with 69B-GAL4 resulted in distal blister phenotype (arrow) with incomplete penetrance (d1 and d3 in a). When DmManf was simultaneously overexpressed, the prevalence of blistered phenotype was significantly decreased (d2 and d4 in a and data not shown). Knockdown of betaTry (betaTrypsin, a serine-type endopeptidase involved in proteolysis) with 69B-GAL4 caused a blistered wing phenotype (arrow) with incomplete penetrance (b1 and b3 in b). The heterozygous DmManf ^Δ96 mutant background significantly suppressed the prevalence of this blistered wing phenotype (b2 and b4 in b and data not shown). The percentages in d1-d4 in A and b1-b4 in B represent the proportion of adult flies with indicated phenotype. Number of analysed adults in (a): d1, 75; d2, 127; d3, 63; d4, 142; and in (b): b1, 325; b2, 189; b3, 286; b4, 236. Scale bar 1 mm. OE, overexpression. c Anti-DmManf (magenta) partially co-localized with mitochondrial marker sqh-EYFP-Mito (green) in the thoracic CNS of 3rd instar larvae. Images consist of four laser confocal sections. Nuclei are shown in blue and gray represents the co-localization of anti-DmManf and sqh-EYFP-Mito. See Additional file 5 for 3D volume rendering. Scale bar 3 μm