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Fig. 1. GFP-PCNA accumulates in the embryonic nucleus during S phase through an
aphidicolin-dependent process. (A) mRNA encoding GFP-PCNA (green, final
concentration 0.0013 µg/µl), together with rhodamine-labelled tubulin
(red, final conc. 0.01 µg/µl) were microinjected 100 minutes before
two-channel confocal image recording. Time after fertilization is indicated.
Second cell cycle, 101-133 minutes; S phase of the third cell cycle, 134-139
minutes. Scale bar: 50 µm. (B) GFP-PCNA protein was microinjected before
fertilization. Confocal images of the nuclei at various times during the first
cell cycle of control embryos (upper panel; scale bar, 15 µm) and embryos
treated with aphidicolin (20 µg/ml at 15 minutes pre-fertilization: lower
panel; scale bar, 100 µm) are shown. The measured average intensity of
fluorescence of the female nucleus throughout the first cell cycle in both
conditions is also shown. The time of each image corresponds to the time
indicated on the graph. Aphidicolin caused a cell cycle block and NEB was not
observed. The graph shows mean±s.e.m. from six control experiments and
from three experiments with aphidicolin treatment. For each experiment a
different female was used.
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