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Fig. 2. XCL100 phosphatase prevents both sperm chromatin decondensation in early
embryos and nuclear accumulation of GFP-PCNA in embryos and Ca2+
ionophore-activated eggs. (A) Wide-field fluorescence microscopy of embryos
microinjected with GFP-PCNA (control) and, in addition, with XCL100 (1.5 µM
and 2.5 µM final concentration) before fertilization and imaged at 65
minutes after fertilization, just before NEB. XCL100 (2.5µM) blocked the
cell cycle and NEB did not subsequently occur. Arrow indicates uncondensed
sperm chromatin. (B) Nuclear GFP-PCNA fluorescence intensity
(mean±s.e.m., control, n=6; 1.5 µM XCL100, n=3;
2.5 µM XCL100, n=4; 2.5 µM XCL100 unfertilized eggs,
n=3) measured during the course of the cell cycle in the experiments
shown in A. Five different females were used in experiments with embryos
(graphs and bars) and three females in the experiments with unfertilized
control eggs. Inset shows that 2.5 µM XCL100 prevented NEB. (C) Confocal
images of nuclear GFP-PCNA in control (n=3) and XCL100-microinjected
eggs (n=4) activated with A23187 (20 µM). The time-course of
GFP-PCNA fluorescence intensity in the nucleus is also shown. NEB was not
observed in the presence of XCL100. Scale bar: 50 µm.
