Real-time PCR was performed, as previously described (63). Briefly, RNA was extracted by using a TRIzol reagent (Invitrogen, 15596-018). cDNA synthesis was generated using a reverse transcription kit (Promega, A3500), according to the manufacturer’s recommendations. qPCR reactions were performed with the Rotor-Gene 6000 (Corbett Research Ltd.) thermocycler. The Maxima SYBR Green/ROX qPCR Master Mix (Thermo Fisher Scientific, K0253) was used to produce fluorescently labeled PCR products during repetitive cycling of the amplification reaction, and the melting curve protocol was used to check for probe specificity, as described previously (30). The following primer sets for all amplicons were designed using the Primer-Express 1.0 Software System (Roche): mouse MuRF1 forward (5′-CCAAGGAAAGAGCAGTATGG-3′) and reverse (5′-GCAGCTCTCTGGGTTATTG-3′), mouse p62 forward (5′-TGAAACATGGACACTTTGGCTGGC-3′) and reverse (5′-ACATTGGGATCTTCTGGTGGAGCA-3′), mouse NBR1 forward (5′-GAGATGAGAGGGAGGAGATT-3′) and reverse (5′-CTTCAGAGGAAGCAGAAGAC-3′), mouse GAPDH forward (5′-TTCAACGGCACAGTCAAG-3′) and reverse (5′-CCAGTAGACTCCACGACATA-3′), human MYOD forward (5′-CACAACGGACGACTTCTATG-3′) and MYOD reverse (5′-GTGCTCTTCGGGTTTCAG-3′), human MYOG forward (5′-GCGTGTAAGGTGTGTAAGAG-3′) and MYOG reverse (5′-GCCTCATTCACCTTCTTGAG-3′), and human GAPDH forward (5′-CGCTTCGCTCTCTGCTCCT-3′) and reverse (5′-CCGTTGACTCCGACCTTCAC-3′).

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