Localization of Macf1a in stage I oocytes.

A-C) Macf1a immunostaining (green) in stage I oocytes shows localization nuclearly and within the Bb (A), and later at the cortex during Bb disassembly (B, C). D) Macf1a staining was negative in a macf1a^sa12708 mutant oocyte. E-G) GFP staining in Gt(macf1a–citrine)^ct68a line also showed Macf1a-Citrine localization to the Bb and at the cortex during disassembly. DAPI (blue/magenta) stains DNA. A-D) N≥ 3 ovaries, 20 isolated WT oocytes were examined (see methods), 16 oocytes showed Macf1 nuclear staining; 18 macf1a^sa12708 oocytes were examined. E-H) N = 3 ovaries, 9 of 20 Gt(macf1a–citrine)^ct68a/+ oocytes showed nuclear staining (H). Representative images from 3 experiments. Dotted white lines outline the nucleus. Images are a sum of 3 single optical confocal sections. Arrowheads indicate the Bb. Scale bars: 20 μm.