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. 2005 May 23;169(4):657–667. doi: 10.1083/jcb.200411034

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Copyright © 2005, The Rockefeller University Press

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Figure 7. — Phosphorylation of IP₃R1 upon application of BK. (A) BK-induced phosphorylation of IP₃R1 occurs in a PKA-dependent manner. (Top) PC12 cells were treated with vehicle or BK (1 μM, 5 min) in the presence or absence of H89 (10 μM) as indicated. Lysates were immunoprecipitated with IP₃R1 antibody, and Western blot analysis of the precipitates was performed by using an antibody designed to recognize substrates phosphorylated by PKA. (Bottom) PC12 cells treated with vehicle (NT) or BK were immunoprecipitated with the phospho-(serine/threonine) PKA substrate antibody, and Western blot analysis of the precipitates was performed by using an antibody against IP₃R1. (B) Time course of the BK induced phosphorylation of IP₃R1. 1 μM BK was applied for the indicated time periods and cell lysates were subjected to immunoprecipitation with antibodies against IP₃R1. Phosphorylation of IP₃R1 was detected as described in A.