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. Author manuscript; available in PMC: 2010 Jun 29.
Published in final edited form as: J Muscle Res Cell Motil. 2009 Apr 21;30(1-2):67–72. doi: 10.1007/s10974-009-9176-y

Figure 1. Cardiac lystaes contain cTnT kinase activity that is blocked by GW5074 (a Raf inhibitor) and not by inhibitors of various other serine/threonine kinases.

Figure 1

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.