Additional file 3: of In vivo screening reveals interactions between Drosophila Manf and genes involved in the mitochondria and the ubiquinone synthesis pathway

Examples of candidate genes for interacting partners of DmManf. A tiff file. A) MS1096-GAL4 was used to silence candidate genes in wild type, heterozygous DmManf Δ96 mutant (DmManf Δ96 /+) and DmManf overexpression (+ DmManf OE) backgrounds. With many candidate genes, both heterozygous DmManf Δ96 mutant background and DmManf overexpression resulted in more severe phenotype. For example, knockdown of Tom20 (Translocase of outer membrane 20, a mitochondrial transmembrane transporter protein) showed a mildly wrinkled wing phenotype in wild type background (a1). When heterozygous DmManf Δ96 mutant background (a2) or simultaneous DmManf overexpression was used (a3), wings were strongly wrinkled. Similarly, knockdown of Cdk12 (a cyclin-dependent protein serine/threonine kinase; b1-b3) showed stronger phenotype both in heterozygous DmManf Δ96 mutant and DmManf-overexpressing backgrounds. Complete list of the alterations is presented in Fig. 4. B) Quantitative analysis of ubiquitous knockdown of candidate genes CSN3, Irbp, COQ7 and ubiquinone synthesis related gene COQ2 showed altered pupal viability with DmManf overexpression (+ DmManf OE, red) in comparison to wild type background (− DmManf OE, blue). tub-GAL4/+ flies were used as wild type control. C) Proportion of emerged adults when candidate genes were ubiquitously knocked down with tub-GAL4 with (+ OE) or without (− OE) DmManf overexpression. *, P < 0.05; ***, P < 0.001, Student’s t-test. Amount of pupae and adults analysed in B-C are presented in Additional file 4. Proportion of Tb+ pupae was normalized to experimentally determined proportion of Tb+ pupae (see Additional file 8, wild type and wild type/SM6-TM6). (TIFF 1442 kb)