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Fig. 2. Dynamics of ERK phosphorylation in primary hepatocytes (A), RTgill-W1 cells
(B) and RTH-149 hepatoma cells (C) from rainbow trout exposed to
hypo-osmolarity, 30 nmol l–1 EGF or 10 µmol
l–1 CuCl2, for up to 60 min as indicated. Numbers
below the blots are changes in pERK abundance, expressed relative to controls
at time zero, denoted as means ± s.e.m. of at least 3 independent
experiments. *Statistically significant difference from the control value, as
assessed on non-normalized data (P<0.05). c60', controls
incubated in standard conditions for 60 min. Activated ERK was determined by
western blot analysis using an antibody against the dually phosphorylated form
of ERK. The main band detected with this antibody migrated at approximately 44
kDa. In A, the second panel shows an example of a western blot analysis of
total ERK abundance using an antibody against the phosphorylated and
non-phosphorylated form of ERK (tERK). Similar analyses were made for all cell
types under all conditions and indicated that ERK abundance was not altered by
the treatments over the time course studied.
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