The transmembrane action potentials were then recorded by several main-stream glass microelectrodes located in a distance of 200 um Dasatinib BMS-354825 from one another. Recording of the motion potentials The hearts, excised quickly from your animal, were mounted on a Langendorff apparatus and were then perfused with welloxygenated normal Krebs solution at a consistent pressure. After 10 to 15 min of stabilization, the monophasic action potentials were recorded with suction electrodes positioned on the top of either the right or left ventricle, which showed spontaneous beating at 200 beats/min to 300 beats/min. Immunoblotting and immunohistochemistry After 10 to 15 min of stabilization, the hearts mounted on the Langendorff apparatus were perfused with Krebs solution, including reagents. At the beginning of fibrillation, at a higher level stage of fibrillation and after treatment Papillary thyroid cancer with reagents, the ventricular tissue specimens were removed from the Langendorff apparatus and subjected to Western blotting and immunohistochemistry. The tissue samples were frozen in liquid nitrogen for Western blotting or were immersed in repairing solutions for immunohistochemistry. The phosphorylation of Cx43 was examined by Western blotting applying anti mouse immunoglobulin G as a primary antibody and mouse monoclonal anti Cx43 antibody as a secondary antibody, or the rabbit polyclonal anti Cx43 antibody as a primary antibody and anti rabbit IgG as a secondary antibody. The total number of Cx43 was assessed by the mean density of the total band found using the mouse monoclonal anti Cx43 antibody as a primary antibody and anti mouse IgG as another antibody. The methods for Western blotting and the examination of the phosphorylation of Cx43 have been previously described. For that immunohistochemistry of Cx43, the rabbit polyclonal anti Cx43 antibody was used as a primary antibody, and goat anti rabbit IgG was used as a secondary antibody. The expression of immunoreactive ALK inhibitor areas of Cx43 in the intercalated disk and localization of Cx43 were examined by immunohistochemistry. Immunofluorescence was detected by confocal laser scan microscopy on the preparations sliced to 10 um in thickness. The procedures for analysis and immunohistochemistry of immunofluorescence have been previously described. Measurement of tissue angiotensin II The expression of angiotensin II in cardiac tissue was examined by Western blotting utilizing rabbit anti IgG and AII. Identification of isoforms of protein kinase C The identification of protein kinase C isoforms was done by Western blotting using a polyclonal antibody for PKC B1, PKC, PKC B2, PKC, PKC  and PKC ..