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Fig. 2. Characterization of complexin II-knockdown cells. (A) Western blot analysis for complexin II, syntaxins 3, syntaxin 4 and synaptotagmin II in four knockdown clones (kd-1 to kd-4). Expression of complexin II was reduced in knockdown cells, while expression of syntaxins and synaptotagmin II was not affected. (B) Intracellular distribution of complexin II in a kd cell. The fluorescence image was obtained as described in Fig. 1C. (C,D) Timecourse of the intracellular Ca²⁺ concentration in wild-type (C) and kd cells (D), respectively. Cells were sensitized with IgE and loaded with Fura2-AM, and stimulated with antigen (DNP-BSA) at the time indicated by an arrow. Average fluorescence intensity ratios (F340/F360) were plotted against time. Values are obtained from four independent preparations of wild-type cells and four independent knockdown clones [mean±s.e.m. (n=4)]. No significant differences in the Ca²⁺ response were detected between wild-type and kd cells.

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