213286.fig.004a
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213286.fig.004b
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213286.fig.004c
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213286.fig.004d
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213286.fig.004e
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Figure 4: Ethanol impairs insulin-stimulated phosphorylation of ERK MAPK and Akt. Control and ethanol-exposed (50 mM x 48 h) rat cerebellar neuron cultures, seeded into 96-well plates, were stimulated with 10 nM insulin for up to 30 minutes. Cells were fixed and used to measure (a) pERK, (b) pAkt, (c) total ERK, and (d) total Akt by cellular ELISA, and results were normalized to H33342 fluorescence. The ratios of (e) pERK/total ERK and (f) pAkt/total Akt were calculated to show relative time-dependent shifts in insulin-stimulated signaling. Graphs depict the mean ± S.E.M. of results generated from 8 replicate assays. Inter-group differences with respect to pERK, pAkt, pERK/Total ERK, and pAkt/Total Akt are statistically significant by area-under-curve analysis.