Fig 6.

Insulin-induced interaction of phospho-ERK with cav-2 distinctively from IGF-1. Hirc-B cells were treated with the indicated nanomolar concentration of insulin or IGF-1 for 10 min. (A) WCL were subjected to immunoblot analysis using anti-phospho-ERK, anti-ERK, anti-cav-2, anti-cav-1 and anti-actin antibodies. The densitometry ratios of phospho-ERK to total ERK were illustrated. The results represent the mean ± S.E. of three independent experiments. (B) WCL were immunoprecipitated with anti-cav-2 antibody and subjected to immunoblot analysis with anti-phospho-ERK, anti-ERK, anti-cav-2, anti-pY19-cav-2 and anti-cav-1 antibodies as indicated. Shown is a representative experiment that was repeated three times. (C) COS-7 cells were treated with or without IGF-1 (10 nM) or insulin (100 nM) for 10 min. Equal amount of WCL were immunoprecipitated with anti-cav-2 antibody and subjected to immunoblot analysis with anti-phospho-ERK, anti-ERK, anti-cav-2, anti-pY19-cav-2 and anti-cav-1 antibodies as indicated. Shown is a representative experiment that was repeated three times.