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Fig. 4. CaM interacts with MARCKS in vitro and in vivo in a Ca²⁺-dependent manner. (A, left) Total protein on PVDF membrane stained with Naphthol Blue Black before exposure to overlay buffer. (A, right) A parallel lane immunostained to locate MARCKS. (A, middle) CaM overlay. The lanes were treated as follows: 1, no Ca²⁺ was added to the overlay buffer and 10 mM EGTA was included; 2, overlay buffer contained 2.5 mM CaCl₂. (B) Two freshly dissociated cells labelled for both CaM and MARCKS. (top) Cell fixed in PSS containing 1 mM Ca²⁺. (bottom) Cell fixed in Ca²⁺-free PSS containing 10 mM EGTA. Arrowheads denote the position of the nucleus.

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