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. 2003 Jun 15;23(12):4984–4995. doi: 10.1523/JNEUROSCI.23-12-04984.2003

Figure 5.

Figure 5.

Analyses of ICI182,780-mediated activation of ERK1/2. A, ICI182, 780 blockade of E2-mediated MAPK stimulation; Western blot analyses of 17β-E2-induced ERK1/2 phosphorylation in cell lysates from cultures pretreated for 30 min with 1 μm ICI182,780. This increased duration of ICI182,789 pretreatment abrogates estradiol-stimulated ERK1/2 phosphorylation. B, ICI182,780 dose–response analysis. Shown are representative Western blot results of the dose dependency of rapid ICI182, 780-induced ERK1/2 phosphorylation. In primary cultures of cerebellar granule cells treated for 10 min with increasing concentrations of ICI182, 780, rapid increases in ERK1/2 phosphorylation are observed with concentrations between 10-9 and 10-7m, indicating that ICI182,780 is a highly potent agonist of rapid ERK1/2 phosphorylation. C, D, Time course analysis of ERK1/2 phosphorylation stimulated by 10-8m ICI182,780 revealed that ERK1/2 phosphorylation was increased significantly at 10 and 15 min after the addition of 10-8m ICI182,780, and with longer exposure ERK1/2 phosphorylation rapidly returned to baseline levels. For each experiment the results are normalized to total ERK-IR and are the mean fold induction ± SEM of six different experiments. *,***See Materials and Methods.