Figure 3

cAMP enahnaces the IR-induced phosphorylation of IKKβ and nuclear translocation of p65. (A) Reh cells were treated with or without forskolin (80 μM) for 30 min before irradiation (10 Gy). Cells were harvested at the indicated times and subjected to Western blot analysis with the indicated antibodies. The figure shows 1 representative blot of 4 experiments. (B) Reh cells were treated as in A, harvested at the indicated times and subjected to immunoblot analysis with the indicated antibodies. The figure shows 1 representative blot of 4 experiments. (C) Reh cells were treated as in A. One half of each sample was used for preparation of total cell lysate and the other half was subjected to cellular fractionation to obtain nuclear lysate. The lysates were then analyzed by immunoblotting with the indicated antibodies. The figure shows 1 representative experiment of 3. (D) Reh cells were cultured in the absence or presence of Bay 11-7082 (5 μM) for 90 min before treatment with or without forskolin (80 μM) for 30 min. Cells were then exposed to 10 Gy IR and harvested after 2 h. One half of each sample was used for preparation of total cell lysate and the other half was subjected to cellular fractionation to obtain nuclear lysate. The lysates were then analyzed by immunoblotting with the indicated antibodies. The figure shows 1 representative experiment of 3.