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Fig. 3. PLC activation involves positive feedback from intracellular Ca²⁺ mobilization. (A) Simultaneous evanescent wave microscopy fluorescence recording of cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i) with Fura Red (red trace) and PLC activity with PH_PLC-GFP (green trace) in a MIN6 cell stimulated with 100 µM carbachol in Ca²⁺-deficient medium (0 Ca²⁺/2 mM EGTA). The Fura Red trace has been inverted to show [Ca²⁺]_i increases as upward deflections. (B) Carbachol-induced PH_PLC-GFP translocation and [Ca²⁺]_i response in a cell preincubated for 40 minutes with 1 mM BAPTA acetoxymethyl ester and maintained in Ca²⁺-deficient medium from 5 minutes prior to stimulation. (C) PH_PLC-GFP translocation and [Ca²⁺]_i response in a cell maintained in Ca²⁺-deficient medium containing 100 µM cyclopiazonic acid. (D,E) Mean±s.e.m. for peak PH_PLC-GFP translocation and [Ca²⁺]_i responses. ^*P<0.01; ^**P<0.001.

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