Chromocenter Disruption in Drosophila Testes

All fly stocks were raised on standard Bloomington medium at 25°C. The following fly stocks were used: D1^EY05004(BDSC17340), Df(3R)BSC666 (BDSC26518), UAS-Omi^RNAi (BDSC55165), UAS-GFP-nls (BDSC4776) and UAS-GFP-ER-SR (BDSC59042) were obtained from the Bloomington Drosophila stock center. D1^LL03310 (DGRC140754) and Df31-GFP (DGRC110806) were obtained from the Kyoto stock center. nos-gal4 (Van Doren et al., 1998 (link)), hs-flp;nos-FRT-stop-FRT-gal4,UAS-GFP (Salzmann et al., 2013 (link)), UAS-H2A-YFP (Bellaïche et al., 2001 (link)) and B1 LacO (Vazquez et al., 2002 (link)) have been previously described. A stock containing B chromosomes, mtrm¹²⁶ +B (Bauerly et al., 2014 (link)), was a kind gift from Scott Hawley. Chromocenter disruption was scored in Drosophila testes by assessing {AATAT}_n morphology in GFP +cells that were generated as follows in control (hs-flp; nos-FRT-stop-FRT-gal4,UAS-GFP) and D1 mutant (hs-flp;nos-FRT-stop-FRT-gal4, UAS-GFP;D1^LL03310/Df) flies. Testes were dissected 24 hr following a 20 min heat shock at 37°C. Chromocenters were considered disrupted in Drosophila and mouse when satellite DNA adopted thread-like morphology in interphase nuclei. Micronuclei were scored in 0-3 d testes where early germ cell chromosomes were labeled with H2A-YFP. The genotypes used were, control – nos > H2 A-YFP and D1 mutant – nos > H2 A-YFP; D1LL03310/Df.