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Fig. 7. Stimulus-specific immunoprecipitation and photoaffinity cross-linking of MARCKS and MARCKS ED peptide with CaM. (A) Immunoprecipitation of portal-vein homogenates with an anti-CaM antibody followed by immunoblot with an anti MARCKS antibody (left) or anti-CaM antibody (right) after exposure of muscles to: no stimulus; 2.5 minutes KCl; or 2.5 minutes DPBA. A typical western blot is shown above the graph of mean±s.e. densitometry for repeated experiments. *, P<0.05; **, P<0.01 compared with unstimulated cells; ###, P<0.001 compared with KCl treatment. n=4. (B) Coomassie-stained SDS-PAGE gel of CaM mixed with ED-BPM peptide and exposed to no light (lane 1), UV light in the absence of Ca (lane 2) and UV light in the presence of Ca²⁺ (lane 3). (C) CaM immunoblot of portal-vein homogenates after muscles were chemically loaded with ED-BPM peptide and exposed to UV light in the presence of no stimulus (lane 2), 51 mM KCl PSS (lane 3) or DPBA (lane 4). Also, as a control, a mock preparation not loaded with peptide but exposed to UV light is shown (lane 1). (D) Average densitometry of the upper band in three experiments performed as described in C. #, P<0.05; ###, P<0.001 compared with unstimulated cells; ***, P<0.001 compared with DPBA treatment.

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