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Fig. 1 | BMC Genetics

Fig. 1

From: In vivo screening reveals interactions between Drosophila Manf and genes involved in the mitochondria and the ubiquinone synthesis pathway

Fig. 1

Knockdown of DmManf by UAS-DmManf-RNAi is effective and specific. a-b Quantification of pupae (a) and adults (b) of ubiquitous knockdown of DmManf with tub-GAL4 driver. Ubiquitous knockdown of DmManf was lethal at larval stage with few escapers to pupal stage. UAS-mCD8-GFP was used as a dose control for UAS/GAL4 binary expression system, tub-GAL4/+ as a wild type control. Amount of pupae and adults analysed are presented in Additional file 8. Proportion of Tb+ pupae was normalized to experimentally determined proportion of Tb+ pupae (see Additional file 8, wild type and wild type/SM6-TM6). c-d Quantitative RT-PCR (c) and Western blot (d) analyses revealed that ubiquitous knockdown of DmManf resulted in robustly reduced DmManf mRNA and protein levels in larvae collected 50–54 h after egg laying. Homozygous DmManf Δ96 mutant showed diminished level of maternally contributed DmManf mRNA and protein. tub-GAL4 > UAS-lacZ flies were used as a wild type control. DmManf mRNA levels were normalized to wild type. Alpha-tubulin was used as a loading control for Western blot analysis. e Knockdown of DmManf with en-GAL4 and salm-GAL4 drivers showed a loss of DmManf immunoreactivity (magenta) in GAL4-expressing pattern. UAS-mCD8-GFP was used to detect GAL4 expression (green). Single laser confocal sections. Scale bar 50 μm

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