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. 2004 May 5;24(18):4324–4332. doi: 10.1523/JNEUROSCI.5227-03.2004

Figure 1.

Figure 1.

Neurotrophin-induced CREB phosphorylation is mediated by an ERK1/2-activated kinase. A, B, Cortical neurons were pretreated with 10 μm KN62 or vehicle and stimulated with BDNF or NMDA for the indicated times (in minutes). Protein extracts were immunoblotted for phospho-cAMP response element-binding protein, ERK2, phospho-CaMKIV, or CaMKIV. C, Cortical neurons were pretreated with 10 μm LY294002 or vehicle and stimulated with BDNF for the indicated times (in minutes). Protein extracts were immunoblotted for phospho-cAMP response element-binding protein, phospho-Akt, and total CREB. D, Rat cortical neurons were treated with 50 ng/ml BDNF or vehicle for 5 min. Akt1 was immunoprecipitated and assayed for kinase activity against GST-cAMP response element-binding protein or GST-Bad. The kinase reactions were separated by SDS-PAGE, and the bands corresponding to the substrate were autoradiographed. Similar results were seen in three experiments. E, Cortical neurons were pretreated with 10 μm PD184352 or vehicle and stimulated with BDNF or NT3 for the indicated times (in minutes). Protein extracts were immunoblotted for phospho-CREB, phospho-ERK, phospho-RSK1/2, phospho-Akt, phospho-ERK5, and total ERK2. F,G, Cortical neurons were pretreated with 5 μm SB202190 or vehicle and stimulated with BDNF or sodium arsenite (As) for the indicated times (in minutes). Protein extracts were immunoblotted for phospho-cAMP response element-binding protein, phospho-HSP27, ERK2, and HSP27.