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Fig. 3. ERK inactivation in late two-cell-stage embryos induces developmental
arrest in the G2 phase, just before M phase in the four-cell stage.
(A) BrdU incorporation into the nucleus was investigated. Late
two-cell-stage embryos were cultured in the presence of BrdU and U0126 or BrdU
and DMSO (control), fixed, and stained with anti-BrdU antibody. Fluorescence
was viewed with a confocal microscope at the early and late four-cell stages.
(B) Cyclin B1 (cyc B1) accumulation in the nucleus and phosphorylation
of histone H3 (p-Histone H3) were examined. Late two-cell-stage embryos were
cultured in the presence of U0126 or DMSO (control). Embryos were fixed and
stained with anti-cyclin B1, Hoechst or anti-phospho-histone H3 antibody.
Fluorescence was viewed with a confocal microscope (cyc B1) or a DeltaVision
Image Restoration Microscope (Hoechst and p-Histone H3). Scale bars: 20
µm.
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