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Mouse lung fibroblasts were serum starved overnight and then stimulated with insulin-like growth factor-I (100 ng/ml) for the indicated times. Cells were lysed, and proteins were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Each membrane was blotted with the indicated phospho-antibody [(A) IGF-1R; (B) IRS-1, IRS-2 (C) AKT, ERK] and then stripped and reblotted with antibody to total protein.
