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. 2013 Apr 9;8(4):e61132. doi: 10.1371/journal.pone.0061132

Figure 4. PMS1077 inhibited TNF-α induced IκB-α degradation, IκB-α phosphorylation, p65 phosphorylation.

Figure 4

(A) TNF-α induced IκB-α degradation, IκB-α phosphorylation. HEK293T cells were treated with TNF-α (20 ng/ml) for the indicated periods of time; (B) PMS1077 inhibited TNF-α induced IκB-α degradation. HEK293T cells were treated with vehicle or PMS1077 (20, 40, 60 and 80 µM) or PMS601 (80 µM) and TPCA-1(1 µM), then incubated with TNF-α (20 ng/ml) for 0.5 h; (C) PMS1077 inhibited TNF-α induced IκB-α phosphorylation. HEK293T cells were treated with vehicle or PMS1077 or PMS601 or TPCA-1 as in (B), then incubated with TNF-α (20 ng/ml) for 2 h; (D) DU145 cells were treated with vehicle or PMS1077 (40 and 80 µM) and TPCA-1(1 µM), then cells were either left untreated or exposed to TNF-α (20 ng/ml) for 12 h. After treatment, the whole cell lysates in (A), (B), (C), and (D) were prepared and analyzed by Western blotting with antibodies for IκB-α, phosphorylation-IκB-α (Ser-32), phosphorylation-p65 (Ser-536) and GAPDH. All data shown are representative of three independent experiments.