Flies were maintained using standard methods. Wild-type stocks were Oregon R (immunohistochemistry) and oskar166 (SCAR antibody controls and actin quantitation) (Lehmann and Nusslein-Volhard, 1986 (link)). See Flybase (http://flybase.bio.indiana.edu ) for details concerning fly stocks. Alleles used were Arp3EP(3)3640 (Rørth, 1996 (link); Berkeley Drosophila Genome Project), Wsp1, Wsp3 (Ben-Yaacov et al., 2001 (link)), Arpc1R337st, Arpc1Q25sd, Arpc1W108R (Hudson and Cooley, 2002 (link)), SCARk13811 (Spradling et al., 1999 (link); Berkeley Drosophila Genome Project), and SCARΔ37.
Germline clones were generated as described (Chou and Perrimon, 1996 (link)) by heat shock of hs-FLP; ovoD FRT40A/SCAR FRT40A larvae, hs-FLP; FRT82B ovoD/FRT82B Wsp larvae, or hs-FLP; ovoD FRT40A/Arpc1 FRT40A larvae. Adult germline clone females were mated to Oregon R males (blastoderm and oogenesis analysis) or to SCARk13811/CyO en-lacZ males (SCARmat/zyg) or Df(3R)3450/TM6B abdA-lacZ males (Wspmat/zyg) (CNS). Mosaic head clones were obtained in ey-FLP; Arpc1Q25sd FRT40A/l(2)cl-L31 FRT40A and ey-FLP; SCARΔ37 FRT40A/l(2)cl-L31 FRT40A flies. SCARk13811 and Wsp3 germline clones (blastoderm), Wsp3 germline clones (CNS), and all zygotic mutants were generated at 25°C. Arpc1R337st germline clones (blastoderm) and SCARk13811 germline clones (CNS) were generated at 20–22°C.
We observed no contribution of zygotic gene activity to the blastoderm defects of embryos derived from SCARk13811 and Arpc1R337st germline clones (unpublished data). Wspmat embryos include embryos defective for both maternal and zygotic Wsp function and embryos defective only for maternal Wsp function.
Germline clones were generated as described (Chou and Perrimon, 1996 (link)) by heat shock of hs-FLP; ovoD FRT40A/SCAR FRT40A larvae, hs-FLP; FRT82B ovoD/FRT82B Wsp larvae, or hs-FLP; ovoD FRT40A/Arpc1 FRT40A larvae. Adult germline clone females were mated to Oregon R males (blastoderm and oogenesis analysis) or to SCARk13811/CyO en-lacZ males (SCARmat/zyg) or Df(3R)3450/TM6B abdA-lacZ males (Wspmat/zyg) (CNS). Mosaic head clones were obtained in ey-FLP; Arpc1Q25sd FRT40A/l(2)cl-L31 FRT40A and ey-FLP; SCARΔ37 FRT40A/l(2)cl-L31 FRT40A flies. SCARk13811 and Wsp3 germline clones (blastoderm), Wsp3 germline clones (CNS), and all zygotic mutants were generated at 25°C. Arpc1R337st germline clones (blastoderm) and SCARk13811 germline clones (CNS) were generated at 20–22°C.
We observed no contribution of zygotic gene activity to the blastoderm defects of embryos derived from SCARk13811 and Arpc1R337st germline clones (unpublished data). Wspmat embryos include embryos defective for both maternal and zygotic Wsp function and embryos defective only for maternal Wsp function.
