Gene expression analysis of differentiating skeletal muscle myoblast cell line overexpressing MyoD

We tested the hypothesis that ectopic expression of the MyoD-dependent gene program in differentiating myoblasts and myotubes compromises the integrity of the plasma membrane/sarcolemma by using a doxycycline-inducible system to overexpress MyoD in C2C12 myoblasts during differentiation. The overarching goals of this analysis pipeline were to (a) determine changes in transcriptional profiles upon sustained overexpression of MyoD during and upon differentiation; and, (b) utilize these profiles to identify potential drivers of sarcolemmal fragility that exacerbate myofiber damage and loss in dystrophic muscle. We used microarrays to generate information about transcriptional profiles in differentiating myoblasts and myotubes overexpressing the myogenic transcription factor myoblast determination protein 1 (MyoD1). Low passage naïve or Lentiviral-transduced C2C12 cells were subjected to differentiation in 2% horse-serum containing media upon doxycycline (dox)-induced overexpression of MyoD. Total RNA was isolated from cells on third day of differentiation and subjected to gene array analysis