RNA extraction (myoblasts, 48 h-MuRC and 96 h-MuRC) was performed with TRIzol reagent (Life Technologies) according to the manufacturer's protocol. After measuring sample concentrations, 0.5 µg of total RNA was reverse-transcribed with the PrimeScript™ RT reagent Kit (TaKaRa) according to the manufacturer's instructions. qRT-PCR experiments were all performed at the iGE3 Genomics Platform of the University of Geneva (http://www.ige3.unige.ch/genomics-platform.php ) on an SDS 7900 HT instrument (Applied Biosystems). Raw threshold-cycle (Ct) values obtained with SDS 2.2 (Applied Biosystems) were imported into Excel. Normalization factor were calculated using the geNorm method [34 (link)]. Primers used for qRT-PCR were: Hs β2-microglobulin forward, 5′-TGCTCGCGCTACTCTCTCTTT-3′; Hs β2-microglobulin reverse, 5′-TCTGCTGGATGACGTGAGTAAAC-3′; Hs EEF1A1 forward, 5′-AGCAAAAATGACCCACCAATG-3′; Hs EEF1A1 reverse, 5′-GGCCTGGATGGTTCAGGTA-3′; Hs Pax7 forward, 5′- AAACACAGCATCGACGGCA-3′; Hs Pax7 reverse, 5′- CTCGTCCAGCCGGTTCC-3′; Hs MyoD forward, 5′- TGCCACAACGGACGACTTC -3′; Hs MyoD reverse, 5′- CGGGTCCAGGTCTTCCGAA-3′; Hs HES1 forward, 5′- CAGATGACGGCTGCGCT -3′; Hs HES1 reverse, 5′- TCGGTACTTCCCCAGCACAC-3′.
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