SRS116607 SAMN00115074 7227 Drosophila melanogaster Immunoprecipitation was performed using anti-Aub and anti-AGO3 (polyclonal) antibodies immobilized on Dynabeads Protein G (Invitrogen). Immunoprecipitation buffer contained 50 mM Tris-HCl (pH8.0), 150 mM NaCl, 1 mM dithiothreitol (DTT), 0.1% SDS, 1% Triton-X, 0.5% sodium deoxycholate, 2µg/ml pepstatin, 2µg/ml leupeptin, and 0.5% aprotinin. Approximately 1500 testes were used per immunoprecipitation. The reaction mixtures were rocked at 4*C for 1 h, the bead fractions were washed five times with binding buffer and the protein pools bound to the beads were eluted with SDS-PAGE sample buffer. After heating to 95*C for 5 min, protein samples were run on SDS-PAGE gels and processed for western blot analysis. SRS116608 SAMN00115075 7227 Drosophila melanogaster Immunoprecipitation was performed using anti-Aub and anti-AGO3 (polyclonal) antibodies immobilized on Dynabeads Protein G (Invitrogen). Immunoprecipitation buffer contained 50 mM Tris-HCl (pH8.0), 150 mM NaCl, 1 mM dithiothreitol (DTT), 0.1% SDS, 1% Triton-X, 0.5% sodium deoxycholate, 2µg/ml pepstatin, 2µg/ml leupeptin, and 0.5% aprotinin. Approximately 1500 testes were used per immunoprecipitation. The reaction mixtures were rocked at 4*C for 1 h, the bead fractions were washed five times with binding buffer and the protein pools bound to the beads were eluted with SDS-PAGE sample buffer. After heating to 95*C for 5 min, protein samples were run on SDS-PAGE gels and processed for western blot analysis.