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. 2011 Apr 20;6(4):e18646. doi: 10.1371/journal.pone.0018646

Figure 3. Sustained activation of JNK after TNFα plus ActD is ROS-dependent and involved in caspase-3/-7 activation.

Figure 3

(A) Western blot analysis of JNK and phospho-JNK (P-JNK) after treatment with TNFα (25 ng/ml) for the indicated times. (B) JNK and P-JNK levels determined by Western blot analysis in cells treated with TNFα (25 ng/ml) and ActD (ActD) (0.4 µg/ml) for the indicated times. (C) Analysis of JNK and P-JNK by Western blot after treatment of hepatocytes with TNFα+ActD with or without preincubation with BHA 100 µM for the indicated times. Actin is used as loading control. One representative experiment is shown. (D) Caspase-3/-7 activation of cells treated with TNFα+ActD with or without preincubation with the JNK inhibitor SP600125 20 µM. For comparison, cells were also treated with FasL (50 ng/ml) for 6 hours with or without costimulation with TNFα+ActD for the indicated times. Means of at least three independent experiments ± s.d. are shown (# P<0.05 versus TNFα+ActD-treated cells, Student's t-test).