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. 2006 Jun;26(12):4586–4600. doi: 10.1128/MCB.01422-05

FIG. 6.

FIG. 6.

Treatment of B82L cells with 8CPT-cAMP activates Erk1/2 as well as RSK1 and dissociates RSK1 from RI. (A) B82L cells (250,000 cells/35 mm dish) were treated with 8CPT-cAMP (100 μM) for the indicated times. Proteins (30 μg) were immunoblotted for phospho-RSK1, phospho-Erk1/2, total RSK1, and total Erk1/2. The last two served as loading controls. Results representing two similar experiments are shown. (B) B82L cells (1 × 106 cells/100 mm dish) were treated with or without 8CPT-cAMP (100 μM) for 30 min. The RI was then immunoprecipitated from cell lysates (500 μg protein), and the presence of RSK1 in the IPs was monitored by immunoblotting. Mouse IgG2b (mIgG) was used in control IPs. Results representing three experiments are shown. (C) B82L cell lysates (500 μg protein) were treated with or without 8-CPT cAMP (100 μM) for 30 min. PKAc was then IP. RSK1 and RI in the IPs were monitored by immunoblotting. Rabbit IgG (IgG) was used in control IPs. Results representing three experiments is shown.