Figure 2. CycB triggers reorganization of the cytoskeleton. (A–C) RLC-GFP (A), GFP-moesin (B; see Video 2, available), and GFP-Nuf (C; see Video 3) embryos were injected with CycB at early interphase of cycle 13. The schematic describes the injection and imaging sequence. The timing of CycB injection in GFP-Nuf was performed at a time when Nuf was concentrated at the MTOC (midinterphase). Arrowheads mark the sites of injection. Top row insets are enlarged images of the control area, which is distant from the site of injection (white boxes), and bottom row insets are enlarged images of the CycB-injected area (blue boxes). The RLC-GFP, GFP-moesin, and GFP-Nuf embryos are representative of 15, 4, and 3 injected embryos, respectively. NEB is determined when the GFP markers fill the nucleoplasm. The blue and orange outlines mark the areas where CycB has an effect on the cytoplasm and the nucleus, respectively. Time is given in minutes/seconds. Bars: (panels) 20 μm; (insets) 10 μm. (D) The timing of cytoplasmic and nuclear CDK1 activation after CycB injection was monitored with RLC-GFP embryos. CycB was injected at 65 or 32 μM at interphase onset, and the embryo was examined within 30 s after injection. The timing of cytoplasmic CDK1 activation was determined when RLC-GFP disappeared from the focal plane. The timing of nuclear CDK1 activation was determined when RLC-GFP filled the nucleoplasm (NEB). For each embryo, we monitored the timing of cytoplasmic and nuclear CDK1 activation in control and injected areas. Data are represented as mean ± AVD. n, number of embryos injected.
Figure 2.

CycB triggers reorganization of the cytoskeleton. (A–C) RLC-GFP (A), GFP-moesin (B; see Video 2), and GFP-Nuf (C; see Video 3) embryos were injected with CycB at early interphase of cycle 13. The schematic describes the injection and imaging sequence. The timing of CycB injection in GFP-Nuf was performed at a time when Nuf was concentrated at the MTOC (midinterphase). Arrowheads mark the sites of injection. Top row insets are enlarged images of the control area, which is distant from the site of injection (white boxes), and bottom row insets are enlarged images of the CycB-injected area (blue boxes). The RLC-GFP, GFP-moesin, and GFP-Nuf embryos are representative of 15, 4, and 3 injected embryos, respectively. NEB is determined when the GFP markers fill the nucleoplasm. The blue and orange outlines mark the areas where CycB has an effect on the cytoplasm and the nucleus, respectively. Time is given in minutes/seconds. Bars: (panels) 20 μm; (insets) 10 μm. (D) The timing of cytoplasmic and nuclear CDK1 activation after CycB injection was monitored with RLC-GFP embryos. CycB was injected at 65 or 32 μM at interphase onset, and the embryo was examined within 30 s after injection. The timing of cytoplasmic CDK1 activation was determined when RLC-GFP disappeared from the focal plane. The timing of nuclear CDK1 activation was determined when RLC-GFP filled the nucleoplasm (NEB). For each embryo, we monitored the timing of cytoplasmic and nuclear CDK1 activation in control and injected areas. Data are represented as mean ± AVD. n, number of embryos injected.

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