Drosophila Transgene Expression Protocols

Drosophila melanogaster flies were raised on standard media; stocks were kept at room temperature and crosses were reared at 25°C or 29°C as indicated. Four Gal4 fly lines were used to induce transgene expression: dpp-Gal4, UAS-GFP/TM6B (Swarup et al., 2015 (link)), Dll-lacZ/Cyo; Hh-Gal4/TM6B (Hall et al., 2017 (link)), tj-Gal4/tj-Gal4 (a gift from Dr Nicholas Harden, Simon Fraser University; Vlachos et al., 2015 (link)), and apterous (ap)-Gal4 (Bloomington Drosophila Stock Center, 3041). Additional stocks used were: puc-LacZ (Martin-Blanco et al., 1998 (link)) and UAS-dsh (Bloomington Drosophila Stock Center, 9453). As controls, the Gal4 drivers were crossed with w¹¹¹⁸ or UAS-GFP flies. To generate transgenic UAS stock lines, patient variant Flag-tagged DVL1 constructs were sent for integration into the attP40 locus on the second chromosome for generation of stably integrated fly strains (BestGene, CA, USA).