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. 2013 Jun 17;8(6):e67633. doi: 10.1371/journal.pone.0067633

Figure 2. Over-expression of WWP1 inhibits TNF-α and IL-6 production induced by LPS, but not by TNF-α, through its C-terminal catalytic activity.

Figure 2

(A) Over-expression of WWP1 and C890A in RAW164.7 stable cell lines. Western blots were used to analyze the His-WWP1 and His-C890A expression in RAW264.7 cells stably transfected with expression plasmid using the His antibody. (B) Effects of over-expression of wild type WWP1 and C890A on LPS- or TNF-α-induced TNF-α and IL-6 secretion. The RAW264.7 stable cell lines of WWP1 and C890A (4×105) were stimulated by LPS (at a final concentration of 400 ng/mL) or TNF-α (at a final concentration of 10 ng/mL) for 12 h. ELISA analysis was used to detect the levels of TNF-α and IL-6 secretion. (C) Effects of over-expression of wild type WWP1 and C890A on LPS- or TNF-α-induced TNF-α and IL-6 transcription. The RAW264.7 stable cell lines of WWP1 and C890A (4×105) were stimulated by LPS (at a final concentration of 400 ng/mL) or TNF-α (at a final concentration of 10 ng/mL) for 8 h. RT-PCR analysis was used to detect TNF-α and IL-6 transcription. **P<0.01 (t-test). Data are representative of three independent experiments (mean and S.D. of three replicates).