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. 2000 Sep 5;192(5):729–740. doi: 10.1084/jem.192.5.729

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© 2000 The Rockefeller University Press

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graphic file with name JEM000333.f4a.jpg — Akt regulation of the NF-κB pathway. (A) wt BMMCs were transfected with an NF-κB/luc reporter plasmid together with an empty vector or a vector encoding wt, E40K, or K179M Akt. 24 h later, cells were sensitized with IgE overnight. Cells were then stimulated with antigen for the last 8 h before luciferase assays. Representative results out of three transfection experiments are shown. (B) wt BMMCs were cotransfected with NF-κB/luc and either an empty vector, DN IκB-α (IκBαM), or DN IKKα (K44M). Results are representative of two experiments. Stim., stimulation. (C) wt and lyn ⁻ ^/− BMMCs were sensitized by IgE and stimulated with antigen for the indicated amounts of time. Cell lysates were analyzed by immunoblotting with antiphospho–IκB-α (phosphorylated Ser-32) antibody followed by reprobing with anti–IκB-α. Results are representative of two similar experiments. Stim., stimulation.

graphic file with name JEM000333.f4b.jpg — Akt regulation of the NF-κB pathway. (A) wt BMMCs were transfected with an NF-κB/luc reporter plasmid together with an empty vector or a vector encoding wt, E40K, or K179M Akt. 24 h later, cells were sensitized with IgE overnight. Cells were then stimulated with antigen for the last 8 h before luciferase assays. Representative results out of three transfection experiments are shown. (B) wt BMMCs were cotransfected with NF-κB/luc and either an empty vector, DN IκB-α (IκBαM), or DN IKKα (K44M). Results are representative of two experiments. Stim., stimulation. (C) wt and lyn ⁻ ^/− BMMCs were sensitized by IgE and stimulated with antigen for the indicated amounts of time. Cell lysates were analyzed by immunoblotting with antiphospho–IκB-α (phosphorylated Ser-32) antibody followed by reprobing with anti–IκB-α. Results are representative of two similar experiments. Stim., stimulation.