By Q. Ivy Fan, Kathleen M. Vanderpool, Jessica O’Connor (auth.), Elissavet Kardami, Larry Hryshko, Nasrin Mesaeli (eds.)
Cardiac phone biology has come of age.
attractiveness of activated or transformed signaling molecules via particular antibodies, new selective inhibitors, and fluorescent fusion tags are yet the various instruments used to dissect signaling pathways and cross-talk mechanisms which could finally permit rational drug layout. realizing the rules of cardiac hypertrophy in all its complexity continues to be a primary target of cardiac learn. because the development of adenovirally mediated gene move, transfection potency isn't any longer a restricting think about the examine of cardiomyocytes. A proscribing think about contemplating phone transplantion as a technique to fix the broken center is mobile availability on the correct time. Cardiac hole junctions, intercellular verbal exchange channels that permit electric and metabolic coupling and play an immense function in arrhythmogenesis at the moment are understood to be beautiful sensors of cardiac switch.
The reviews during this quantity contain dependent stories that made use of innovative technological advances and plenty of really expert reagents to deal with those issues.
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Extra resources for Cardiac Cell Biology
Hertz 18: 245-255, 1993 6. Bishopric NH, Simpson PC, Ordahl CP: Induction of the skeletal a-actin gene in al-adrenoceptor-mediated hypertrophy of rat cardiac myocytes. J Clin Invest 80: 1194-1199, 1987 7. Waspe LE, Ordahl CP, Simpson PC: The cardiac ~-myosin heavy chain isogene is induced selectively in aI-adrenergic receptorstimulated hypertrophy of cultured rat heart myocytes. J Clin Invest 85: 1206--1214, 1990 8. Kariya K, Karns LR, Simpson PC: Expression of a constitutively activated mutant of the ~-isozyme of protein kinase C in cardiac myocytes stimulates the promoter of the ~-myosin heavy chain isogene.
The medium was changed every 48 h. On days 3, 4,5,6, 7 from plating, cells were trypsinized and counted using a hemocytometer. Results and discussion Transient transfection and stable expression of Cx43 constructs Cardiac myocytes were cultured on collagen-coated covers lips or plastic, in 35 mm dishes, while HeLa cells were grown directly on covers lips or plastic. Transient transfections were done using the calcium phosphate precipitation method with modifications, as described previously .
Subcellular fractionation and analysis by Western blotting The Nuclei-EZ kit (SIGMA) was used to obtain nuclei-enriched and cytosolic fractions from cardiomyocyte or HeLa cell homogenates, according to manufacturer's instructions. Fractions (20 Jlg protein/lane) were analysed by SDS-PAGE, using 12% polyacrylamide gels, transferred subsequently to PVDF membranes and probed with anti-Cx43 polyclonal antibodies as described previously . Cell growth Stable cell lines were grown in 10% FBS in DMEM-hi-glucose medium with G418 until they reached 80% confluence.
Cardiac Cell Biology by Q. Ivy Fan, Kathleen M. Vanderpool, Jessica O’Connor (auth.), Elissavet Kardami, Larry Hryshko, Nasrin Mesaeli (eds.)