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Calcium-dependent release of the complexin block. v- and t-cells were incubated overnight before the addition of 1 unit/ml PI-PLC and 1.8 mm EGTA (t = 0, where indicated) to release the complexin GPI anchor and reduce free calcium in the media to 10 μm. At t = 5 min, free calcium was raised to 200 μm (▪, CPX-I (wild type)-GPI; ○, CPX-I-(26-83)-GPI) or kept constant low at 10 μm (□, CPX-I-(26-83)-GPI). Cells were fixed, and data were quantified as in Fig. 1.
