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Figure 3 Ca²⁺ Leak, Ser²⁸⁰⁸-Phosphorylation Level in RyR2, and FKBP12.6

(A) Representative time courses of Ca²⁺ uptake and the ensuing Ca²⁺ leak from SR vesicles in the presence or absence of 30 µmol/l FK506 and/or 100 µmol/l carvedilol (CV) (left). Summarized data of Ca²⁺ leak in normal, CV-untreated, and CV-treated sarcoplasmic reticulum (SR) vesicles (right). (B) Protein kinase A phosphorylation level of RyR2. (C) The amount of RyR2-bound FKBP12.6. To see the direct acute effect of CV on phosphorylation level of RyR2-pSer²⁸⁰⁸ or amount of RyR2-bound FKBP 12.6 in failing SR vesicles, the SR vesicles were mixed with CV (100 µmol/l) for 30 min and then centrifuged, followed by immunoprecipitation and Western blotting. 4W-pacing = pacing at 250 beats/min for 4 weeks; ATP = adenosine triphosphate; other abbreviations as in Figure 2.

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