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Fig. 3. Expression and [³H]Ins(1,4,5)P₃-binding activity of the putative Ins(1,4,5)P₃-binding domain of PtIP₃R_N. (A) Overexpression of GFP alone (top) and GFP fused to the putative Ins(1,4,5)P₃-binding domain (IP₃BD) of IP₃R_N (bottom) in Paramecium 7S cells. (B) Western blot analysis of immuno-precipitated GFP-IP₃BD fusion protein or GFP alone with GFP-specific Ab. (C) [³H]Ins(1,4,5)P₃-([³H] IP₃) binding assay using agarose beads coupled to protein A either with GFP alone or with GFP-IP₃BD fusion protein. Inhibition of specific [³H]Ins(1,4,5)P₃-binding was measured in the presence of 10 µM cold Ins(1,4,5)P₃. The graph represents one out of five experiments.

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