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Fig. 2. (A) Antisense treatment of cardiac mycoytes knocks down MURF-2 mRNA and protein levels. Cardiac myocytes were treated with MURF-2 antisense or control oligonucleotides and RT-PCR was performed. MURF-2 mRNA transcript levels were significantly decreased in cultures treated with MURF-2 antisense oligonucleotides (lane 2) compared to control oligonucleotides (lane 1), although both GAPDH (lanes 5, 6) and MURF-3 (lanes 3, 4) mRNA transcript levels were unchanged. (B) Lysates of cardiac myocytes treated with control (C) or antisense (A) oligonucleotides were harvested 48 hours after treatment, protein assays were performed to ensure equal loading for SDS-PAGE, and transferred to nitrocellulose membranes. The strips were probed with anti-MURF-2 antibodies, revealing that both the 60 kDa and 55 kDa isoforms were significantly decreased in intensity (>50%) at the protein level in the antisense-treated lysates compared to control myocytes.





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