Fig. 4

GFP-Cfap53 localization during cleavage stages. (A-F) Maximal projections of confocal stacks from Tg(ubi:GFP-cfap53) embryos immunolabelled for GFP, γ-tubulin and DAPI from 30 mpf until 4 hpf. (A) At 10 mpf, male and female pronuclei and the female polar body are visible. Some γ-tubulin foci around the male pronucleus can been observed, while GFP-Cfap53 has a diffuse localization (A, n=7). (B) At 15 mpf, both pronuclei are almost fused and some γ-tubulin can be observed around the nuclei, while GFP-Cfap53 localization is still diffuse (B, n=7). (C) At 30 mpf, chromosomes have aligned along the metaphase plate and strong γ-tubulin staining can be observed on either side of it. At this moment, GFP-Cfap53 also starts to localize to both sides of the metaphase plate, where it colocalizes with γ-tubulin (C, n=12). (D) At 45 mpf, the first cell division has completed and one of the two nuclei is shown. Both GFP-Cfap53 and γ-tubulin are visible in broad domains on both sides of the nucleus during prophase (D, n=8). (E) Single cell of an eight-cell stage blastula with segregating chromosomes during telophase of mitosis. GFP-Cfap53 and γ-tubulin colocalize in a more condensed region at both sites of the division plane (E, n=7). (F) Blastula cell in telophase during mitosis at the sphere stage (4 hpf). GFP-Cfap53 and γ-tubulin colocalize to a confined region on both sides of the division plane (F, n=1). Asterisks indicate polar bodies. Gender symbols indicate male or female pro-nucleus. Scale bars: 20 μm. Schematics of embryonic development are adapted, with permission, from Kimmel et al. (1995).