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. 2004 Aug;24(16):7298–7311. doi: 10.1128/MCB.24.16.7298-7311.2004

FIG. 6.

FIG. 6.

Recombinant PKCδ inhibits SRF DNA binding activity. (A) A preparative digest of PKCδ was performed by incubation for 3 h at 37°C with recombinant caspase 3. A PKCδ Western blot shows the liberation of the PKCδ catalytic fragment (PKCδ-CF). These preparative fractions were used in subsequent phosphorylation analyses with PKCδ. (C) Phosphorylation-specific Western blot and SRE EMSA of in vitro kinase assays with activated PKCδ (lanes 2 to 4), caspase-cleaved PKCδ (lanes 5 to 7), and recombinant casein kinase II (lanes 8 to 10). The kinases were used to phosphorylate SRF(His)6 in a time course of 45 to 180 min at 37°C. The control (lane 1) used SRF(His)6 alone under the same conditions without any kinase. Reaction products were used in SRE EMSAs (B) or in a Western blot with a phosphoserine/threonine-phenylalanine (Phe +1) antibody (C).

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