Drosophila type 2 agar

Additional fly strains included spz (spz/rm7, loss of function point mutation [81 (link)], gift from B. Lazzaro), and Myd88 (BDSC 14091, transposable element insertion). Flies were reared on CMY media: 64.3 g/L cornmeal (Flystuff Genesee Scientific, San Diego CA), 79.7 mL/L molasses (Flystuff Genesee Scientific), 35.9 g/L yeast (Genesee Scientific inactive dry yeast nutritional flakes), 8 g/L agar (Flystuff Genesee Scientific Drosophila type II agar), 15.4 mL of antimicrobial mixture [50 mL phosphoric acid (Thermo Fisher, Waltham MA), 418 mL propionic acid (Thermo Fisher), 532 mL deionized water], and 1g/L tegosept (Genesee Scientific). Flies were reared, mated, and housed after infection, at room temperature (25°C) and were kept on a 12 h light/dark light cycle.

Fly strains include Drosophila melanogaster w¹¹¹⁸ (one strain with Wolbachia, one cured of Wolbachia via tetracycline) and D. melanogaster w^k (one strain with Wolbachia, one cured of Wolbachia via tetracycline). The w^k line was isolated in Karsnäs, Sweden in 1960 (white allele named for location of isolation)⁷⁷ and the w¹¹¹⁸ line was isolated in California and described in 1985 (white allele named for date of isolation)^{76 (link)}. Both were maintained in various labs since their isolation. Flies were reared on CMY media: 64.3 g/L cornmeal (Flystuff Genesee Scientific, San Diego CA), 79.7 mL/L molasses (Flystuff Genesee Scientific), 35.9 g/L yeast (Genesee Scientific inactive dry yeast nutritional flakes), 8 g/L agar (Flystuff Genesee Scientific Drosophila type II agar), 15.4 mL of antimicrobial mixture [50 mL phosphoric acid (Thermo Fisher, Waltham MA), 418 mL propionic acid (Thermo Fisher), 532 mL deionized water], and 1g/L tegosept (Genesee Scientific). Flies were kept at 25°C on a 16h light/8 h dark light cycle.