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Figure 5. Effects of p85 overexpression on phosphorylation of Akt and p70S6K induced by 5-MCDE. Subconfluent (90%) monolayers of Cl41 AP-1 mass1 and Cl41 AP-1 p85 mass1 in 6-well plates were subjected to 1 µM 5-MCDE for various periods of time as indicated. The cells were washed once with ice-cold PBS and extracted with SDS-sample buffer. The cell extracts were separated on polyacrylamide–SDS gels, transferred, and probed with one of the specific antibodies as indicated. The protein band specific bound with primary antibodies was detected by using anti–rabbit IgG-AP-linked and ECF Western blotting system.
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