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. Author manuscript; available in PMC: 2018 Jan 11.
Published in final edited form as: Sci Signal. 2013 May 14;6(275):ra35. doi: 10.1126/scisignal.2003708

Fig. 3. USP25 inhibits TLR4-dependent activation of NF-κB and MAPKs in MEFs.

Fig. 3

(A) USP25 inhibits the generation of proinflammatory cytokines in MEFs stimulated with LPS. WT or Usp25−/− MEFs were stimulated with LPS (1 μg/ml) for 3 hours before real-time reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to determine the relative abundances of the indicated mRNAs. Data are means ± SD from three independent experiments. *P < 0.01 by t test. (B and C) Deficiency in USP25 potentiates LPS-induced activation of NF-κB and MAPKs in MEFs. (B) WT or Usp25−/− MEFs were stimulated with LPS (10 μg/ml), and the degradation of IκBα and phosphorylation of ERK, JNK (c-Jun N-terminal kinase), and p38 were analyzed by Western blotting with antibodies against the indicated proteins. Data are representative of three independent experiments. *P < 0.05; **P < 0.01 by analysis of variance (ANOVA). (C) WT or Usp25 −/− MEFs were stimulated with LPS (10 μg/ml) or poly(I:C) (50 μg/ml). Nuclear extracts were prepared and subjected to EMSA analysis with NF-κB, AP-1, and Oct-1 probes. Data are representative of three independent experiments. Asterisk indicates nonspecific bands.