FIG. 7.

Recruitment of the complex by CRG is independent of CRG phosphorylation by PKA. (A) To test for phosphorylation of CRG and KID-G4 by PKA in the Ag-EMSA system, we incubated 100 fmol of each protein without and with 5 U of PKA under the same conditions used in the Ag-EMSA binding reactions. Samples were separated on an SDS–15% PAGE gel and transferred to a membrane, and a Western blot was performed with a primary antibody specific for Ser-133 phosphorylated CREB. (B) Ten femtomoles of each purified protein was included with 5 fmol of probe and 3 μg of RLNE in Ag-EMSA binding reactions as indicated. Five units of PKA were included where shown.