PKA activation reduces phospho-Thr-75 DARPP-32 even when Cdk5 activity
is inhibited. (a) Slices were incubated in the absence
or presence of roscovitine (Rosc) (10 μM or 50 μM) for 60 min,
followed by the addition of forskolin (FSK) (2 μM) for 5 min. The
amounts of phospho-Thr-75 DARPP-32 were quantified by densitometry, and
the data were normalized to the values obtained with untreated slices.
Data represent means ± SEM for 4–15 experiments.
**, P < 0.01 compared with no
addition; †, P < 0.05, ††,
P < 0.01 compared with forskolin alone; §§,
P < 0.01 compared with roscovitine (10 μM)
alone; ¶, P < 0.05 compared with roscovitine (50
μM) alone, Student's t test. (b)
Homogenates of neostriatal slices treated with forskolin at the
indicated concentrations were subjected to immunoprecipitation (IP)
with either a Cdk5 antibody (C-8) or a normal rabbit IgG. The
immunoprecipitates, containing Cdk5 and its cofactor p35, were used to
phosphorylate histone H1 in the absence or presence of roscovitine.
Results similar to those shown here were obtained in two other
experiments.