Figure 4. Effect of phorbol 12-myristate 13-acetate on expression and phosphorylation of c-jun N-terminal kinase and p38MAPK

Subconjunctival fibroblasts were treated with 10 nM PMA for 5 min. For inhibition experiments, we added bisindolylmaleimide I (BIS), SP600125 or SB203580 to the serum-starved cells 30 min prior to phorbol 12-myristate 13-acetate (PMA) treatment. Western blot analysis was employed to examine expression and phosphorylation of c-jun N-terminal kinase (JNK) and p38MAPK. A: JNK phosphorylation was determined with an anti-phospho-JNK specific antibody. The amount of total JNK was examined by reprobing the membrane with an anti-JNK antibody. Arrows mark the locations where JNK proteins should appear. B: p38MAPK activation was evaluated with an anti-phospho-p38MAPK specific antibody. The amount of total p38MAPK was evaluated with anti-p38MAPK antibody after stripping.