Fly Husbandry and Transgenic Lines

Fly stocks were kept at 25°C on a 12-h light and 12-h dark cycle and fed a standard sugar/yeast/agar diet (Bass et al, 2007 (link)). The light intensity in the fly chambers was around 1,000 lux. The flies were reared at controlled larval densities, and once-mated female flies were used for all experiments unless otherwise stated. The flies were snap-frozen with liquid nitrogen. Dissections were carried out in PBS and tissues either directly analysed or frozen on dry ice.
Transgenic flies were backcrossed into the outbred white Dahomey (wDah) or red Dahomey (rDah) wild-type strain (Grönke et al, 2010 (link)) with the endosymbiont Wolbachia for at least six generations, if necessary. The following transgenic fly lines were used in this study: GMR-Gal4 (Bloomington), UAS-miR-210 (Bejarano et al, 2012 (link)), Sima KO/Tm3Sb (Bloomington #14640), Dgk RNAi (Bloomington #41944), GMR-p35 (Bloomington #5774), Df(1)BSC352 (Bloomington #24376), Apc RNAi (VDRC v51468), Vha55 RNAi (VDRC v46553), Fasn1 RNAi (VDRC v29349) (Dietzl et al, 2007 (link)), UAS-Fasn1 (Garrido et al, 2015 (link)), and UAS-bmm-eGFP (Grönke et al, 2005 (link)).