Lysates from resting cardiomyocytes were subjected to in vitro kinase assays without (lane 1) and with troponin complex (lanes 2-11). The following kinase inhibitors were used in the assays: 50 μM H7, 5 μM GF109203X, 5 μM Go6976, 20 μM H89, 5 μM U0126, 10 μM GW5074, 10 nM Rapamycin, 10 μM Y27632, and 10 μM UCN-01. It is worth noting that UCN-01 specifically blocks PDK-1 and checkpoint kinases at low concentrations, but UCN-01 also inhibits PKC isoforms at the concentrations used in these studies. Autoradiogram and Ponceau staining are depicted in the left panels, with a quantification of the data in the right panel. These results were replicated in two similar experiments. Similar results also were obtained in experiments that used lysates from H2O2-treated cardiomyocytes cultures.