Biomedical Engineering Reference
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Fig. 2. EMSAs in primary striatal culture and in rat striatum.
(A)
Striatal extracts
from two rats treated with saline and two rats treated with amphetamine, i.p. Levels of
the protein(s) binding to the ATF enhancer are unchanged (
see also
ref.
7
for further
experimental details), whereas levels of proteins binding to the AP-1 enhancer are
induced by amphetamine.
(B)
In primary striatal culture, treatment with dopamine also
does not affect binding to the ATF site. However, the addition of a phosphorylation-
specifi c antibody, phosphorylated cAMP response element binding protein (pCREB),
reveals that the state of phosphorylation of the protein bound at the ATF site, CREB, is
altered by treatment with dopamine. Indeed, the activity of the protein binding to the
ATF site is regulated by phosphorylation, whereas the activity of the protein(s) binding
to the AP-1 site is regulated by protein concentration. The specifi city of the binding
was established with various antibodies in supershift experiments, and with a panel of
unlabeled enhancer elements in competition experiments
(7)
.
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