Fig. 3. Effects of midazolam on phosphorylation of p44/p42 mitogen-activated protein (MAP) kinase, stress-activated protein kinase (SAPK)/c-Jun N -terminal kinase (JNK), and p38 MAP kinase in A10 cells. Cultured cells were stimulated by 0.1 mm midazolam for the indicated periods. Cell lysates were subjected to sodium dodecyl sulfate - polyacrylamide gel electrophoresis followed by Western blot analysis using antibodies against phosphospecific p44/p42 MAP kinase, p44/p42 MAP kinase, phosphospecific SAPK/JNK, SAPK/JNK, phosphospecific p38 MAP kinase, or p38 MAP kinase. The bars show quantitative representations of the phosphorylation of MAP kinases obtained from laser densitometric analysis. Each value represents the mean ± SD of triplicate determinations. Similar results were obtained in two other cell preparations. *P is less than 0.05 compared with the value in unstimulated cells.