Drosophila Genetic Manipulation Protocol

The following Drosophila melanogaster lines were used in this study: Oregon-R (Bloomington Drosophila Stock Center (BDSC) #6362), da-Gal4 (BDSC#55851), UAS-GFP RNAi (BDSC#9330), UAS-Ada2b RNAi (National Institute of Genetics, Japan (NIG) #9638R-3), white¹¹¹⁸ (w) mutant (BDSC#3605), hmlΔ-Gal4 UAS-2XEGFP (BDSC#30140), and Ada2b^d272 mutant (a gift from Dr. N. Zsindely, University of Szeged) [24 (link),25 (link)] lines. To knock down Ada2b ubiquitously, the da-Gal4 line was crossed with the UAS-Ada2b RNAi line. The resulting progenies (da-Gal4, UAS-Ada2b RNAi; designated as da > Ada2b RNAi) were used for the experiment. The progenies resulting from the cross of da-Gal4 and UAS-GFP RNAi lines were used as controls (da-Gal4, UAS-GFP RNAi; designated as da > GFP RNAi). To knock down Ada2b in hemocytes, the hmlΔ-Gal4 line was used in a similar way. The Ada2b mutant heterozygotes (Ada2b^d272/+) were obtained by crossing the w line with the w; Ada2b^d272/TM6 line. The siblings (TM6/+) of this cross were used as controls. Flies were reared on standard corn meal medium at 25°C.