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Fig. 3. Immunostaining and quantification of CycB. (A-D) Wild-type embryos were
stained with both anti-CycB (green) and anti-histone H1 (red) antibodies. (A)
An energid is outlined and the average pixel intensity within the energid
(green channel only) was measured. The outline was defined by taking the
shortest distance between yolk granules (black) around the nucleus. Because
the average pixel intensities (total pixel numbers within an area divided by
the area) were compared, the exact outlines were not crucial for the
comparison. (B-D) CycB levels were quantified from three regions (anterior,
middle and posterior) in each embryo, averaging the data from three nuclei in
each region. In this example, the mean value of the average pixel intensity of
the CycB signal was ranked, with the lowest CycB level (74.4±6.3) as 1,
the highest (94.6±3.7) as 3 and the middle (85.0±4.1) as 2. (E)
Summary of the rank sum of different cell-cycle phases at cycle 7. The
asterisks indicate that the regional difference is statistically significant
(P<0.04) based on Friedman's Analysis of Variance by Ranks
(Zar, 1999). Scale bars: 20
µm.
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