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CML-HSA treatment activates MAPK family members ERK1/2 and p38 but not JNK. Samples were taken at the indicated times after 100 μg/ml CML-HSA exposure and subjected to SDS-PAGE and Western blotting analysis. CML-HSA treatment activates MAPK family members ERK1/2 and p38 but not JNK. Samples were taken at the indicated times after 100 μg/ml CML-HSA exposure and subjected to SDS-PAGE and Western blotting analysis. Upper panels: probed with an antibody that specifically recognizes the phosphorylated forms. Lower panels: the same membrane was stripped and reprobed with a pan antibody, showing equal protein loading of all lanes. A: ERK1/2; B: p38; and C: JNK. To test the effect of PD98059 and SB203580, cells were pretreated with either inhibitor for 30 min, followed by CML-HSA exposure for another 20 min, a time point at which the peak activation was seen. Results shown are representative of three independent experiments. Densitometric analysis for kinase activation by CML-HSA compared with the basal level (time 0, set as 1.0) is indicated. Chen-Hsiung Yeh et al. Diabetes 2001;50: ©2001 by American Diabetes Association
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