Immunofluorescent analysis of early Drosophila embryos.

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Immunofluorescent analysis of early Drosophila embryos. Immunofluorescent analysis of early Drosophila embryos. Nuclei were counterstained with DAPI (blue). (A and B) Embryos fixed using a standard technique. (A) Embryos (0–1 hr) of pnutmut1 rescued with FLAG-pnut-WT, FLAG-pnut-T509A/S517A, or FLAG-pnut-T509E/S517E transgenes were stained for Pnut (red). (B) Embryos at the syncytial blastoderm stage (1.5–2 hr) were stained for FLAG (green) and Sep1 (red). Side views at the pseudocleavage furrows are shown. (C and D) Manual devitellinization protocol was used (see Materials and Methods). (C) Embryos (1.5–2 hr) were stained for Pnut (green) and actin (red). Top views at the pseudocleavage furrows are shown. (D) Embryos (1.5–2 hr) were stained for Pnut (green). Side views are shown. Bar, 10 µm. DAPI, 4’,6-diamidino-2-phenylindole; WT, wild-type. Katarina Akhmetova et al. G3 2017;8:27-38 ©2018 by Genetics Society of America