GSI treatment blocks alcohol-dependent Hes1 activation.

<p>C2C12 cells were induced to differentiate for 2 days in control, DM containing either 100 mM ethanol, 1 uM L-685, 458 or both. Mean ± SEM (A) Hes1 (n = 4 independent replicates) and (B) MyoD (n = 2 independent replicates) mRNA transcript levels measured by qRT-PCR relative to untreated controls is shown. Black bars represent control medium treated cultures and grey bars represent cultures treated with medium containing alcohol. GSI treatment significantly reduced Hes1 expression in both control cells and cells exposed to alcohol. Induction of Hes1 by alcohol administration in the absence of GSI treatment was not observed in the presence of L-685, 458 consistent with activation of the Notch pathway by alcohol. L-685, 458 treatment mildly increased MyoD expression with a partial reversal of alcohol mediated repression. Asterisk indicates Student’s t-test <i>p</i>≤0.05 and NS indicates non-significant.