Fig. 7. Inhibition of p47/Eyc disrupts nuclear envelope assembly. Wild-type, eyc^l39 (1.5-2 hours at 25°C), and microinjected syncytial blastoderm embryos triple labeled with phalloidin (red), anti-Lamin (green) and YOYO-1 (blue). Confocal images focus on the cortex of the syncytial blastoderm. (A) In wild-type M phase embryos, Lamin staining shows disassembled nuclear envelope while nuclear vesicles congregate the separated chromosomes. (B) Interphase embryos show Lamin staining of nuclear envelopes and a normal hexagonal array of phalloidin staining. Two examples of eyc^l39 embryos show abnormal Lamin staining pattern. (C) A commonly observed eyc^l39 phenotype shows loss of normal staining pattern for the three markers. (D) An optical section of a mutant embryo in the same focal plane as in B shows a cloudy Lamin staining, possibly owing to the aggregation of Lamin-coated nuclear vesicles. (E-H) Microinjection of wild-type stage 2-3 syncytial blastoderm embryos with pre-immune serum (E,G) or anti-Eyc antiserum (F,H). Equal volumes of serum were microinjected at the posterior (right) end of the embryo. (E,F) Projection views of nine sections, 4 µm apart from the cortex to the embryo core. (E) Embryos injected with pre-immune serum show a normal hexagonal array of cortical nuclei (the blue patch at the posterior is a YOYO-1 crystal). (F) Anti-Eyc antiserum-injected embryo shows fewer nuclei in the posterior region with abnormal nuclear aggregates. (G) An optical section through the embryo shows normal, densely-packed cortical nuclei surrounding embryo in a pre-immune serum injected embryo. (H) A corresponding side view of an embryo injected posteriorly with anti-Eyc shows loosely-distributed, spherical nuclei at the posterior. Scale bar: 20 µm in A-D; 100 µm in E-H.

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