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The next day, the suspensions were centrifuged at 8,500 g for 5 min at 4C, and the supernatants were removed

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Removal of Ca2+ by the addition of EGTA or application of Ca2+-channel blockers, verapamil, diltiazem, and Ni2+, inhibited the BK-induced IP accumulation and Ca2+ mobilization, indicating that Ca2+ influx was required for the BK-induced responses