Supplemental Figure 1
-
Fig. S1. Abolition
of the post-fertilisation peak in ERK1 activity prevents later activation of
ERK1 and Cdk1/cyclin B at the time of NEB in control embryos. (A) The second,
later peak in MAP kinase activity at the time of NEB in control embryos (black
circle) and in embryos treated with U0126 10 minutes before fertilization
(black square); also shown is the percentage of embryos undergoing NEB in
control (white circle) and U0126-treated (white square) embryos. (B) Western
blotting of immunoprecipitated (Promega) active MAP kinase dimers, detected
with a second anti-dual phosphorylated MAP kinase antibody (New England
Biolabs). Controls were sampled at 55 and 110 minutes, just before and at the
peak of ERK1 activity (see A, above); embryos treated with 100 mM U0126
10 minutes before fertilisation (+U0126, 100 minutes) were sampled at 100
minutes after fertilisation; a control immunoprecipitation in the absence of
antibody (-AB) at 55 minutes is also shown. (C) hH1 activity around the time of
NEB in controls (black circle) and in embryos treated 10 minutes before
fertilization with 100 mM U0126 (black square). D) hH1 activity measured
using pull-down of Cdk1/cyclin with p13 beads. Results from control embryos and
those treated with U0126 10 minutes before fertilization (U0125-10)
are shown.
