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Fig. 4. FHL3 siRNA knockdown accelerates myogenin protein expression during differentiation. (A) Lysates from differentiating siRNA-transfected C2C12 cells were immunoblotted for MyoD, myogenin or actin. (B) Myogenin protein levels in siRNA-transfected C2C12 cells were quantified after 24 hours of differentiation by densitometry of immunoblots standardised to actin as a loading control and vector levels arbitrarily set at 1 (n=4, *P<0.05, **P<0.01). Error bars represent ± s.e.m. (C) siRNA-transfected C2C12 cells were differentiated for 24 hours, stained with anti-myogenin antibodies (blue) and propidium iodide (red) and imaged by confocal microscopy. Bar, 20 µm. Yellow arrowheads indicate myogenin-positive nucleus. White arrowheads indicate myogenin-negative nucleus. (D) The mean percentage of nuclei expressing myogenin was quantified following 24 hours of differentiation, a minimum of 320 nuclei were scored per transfected siRNA oligonucleotide species, per time point (n=4, *P<0.05, **P<0.01). Error bars represent ± s.e.m.
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