il\VOLVEMENT OF RETII\OIC ACID II{ - MSpace at the University of ...

Cruz, CA, USA). Marker proteins for apoptosis, Bax and BCL-xl, were also detected
using a rabbit IgG Bax and BCL-xl polyclonal antibodies (Cell Signaling Technology
inc., Beverly, MA, USA). PPAR ô protein levels were detected using a rabbit IgG PPAR
ô polyclonal antibody (Sigma-Aldrich CO, St. Louis MO, USA). Primary antibodies
were detected using an goat anti-rabbit IgG horseradish peroxidase conjugate secondary
antibody @io-Rad, Hercules, CA, USA). Molecular weights of the separated proteins
were determined using a standard (Bio-Rad, Hercules, CA, USA) and biotynilated (Cell
Signaling Technology inc., Beverly, MA, USA) protein ladder molecular weight markers.
Detection of membrane-bound proteins was performed using the BM Chemiluminiscence
(PoD) westem blotting system (Roche Diagnostics GmbH, Manheim, Germany). The
bands were visualized with Flour S-Multilmager MAX system (Bio-Rad, Hercules, CA,
USA) and quantified by image analysis software (Quantity One, Bio-Rad, Hercules, CA,
usA).
I.e.Affymetrix gene-chip probe array analyses
Total RNA was isolated and purified (TriReagent kits, Qiagen, Mississauga,
Ontario, Canada) from the hearts. Two independent samples each from the CONT, ADR
and ADR+PROB groups were sent to the Centre for Applied Genomics (Hospital for
Sick Children, Toronto, Ontario, Canada) for Rat Genome anay (tJ34A) analyses.
ll.In Vítro Studies
II.a.Adult Cardiac Myocvtes Isolation
Ventricular myocytes were isolated from the adult male Sprague Dawley rats
using a procedure described earlier (Kirshenbaum et al. 1992). Sprague-Dawley Rats
(250-3009) were injected with sodium heparin. One hour after the injection of heparin,
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