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. 2018 Jun 27;293(34):13191–13203. doi: 10.1074/jbc.RA118.002649

Figure 4.

Figure 4.

C25-140 effect on proinflammatory signaling and T-cell activation. A, endogenous TRAF6 auto-ubiquitination (Ub) in MEF cells upon IL-1β stimulation is reduced after C25-140 treatment. B, C25-140 impairs IL-1β-induced IκBα phosphorylation. pIκBα levels were densitometrically quantified in relation to β-actin. Error bars, S.D.; n = 3 biological replicates were quantified; unpaired t test (two-tailed); ****, p < 0.0001. C, target gene (ICAM-1 and A20) expression is diminished after IL-1β stimulation and C25-140 treatment; error bars, S.D.; n ≥ 3 biological replicates; unpaired t test (two-tailed). *, p < 0.05; **, p < 0.01; ****, p < 0.0001. D, endogenous TRAF6 auto-ubiquitination after P/I stimulation is reduced after C25-140 treatment. E, C25-140 decreases IκBα phosphorylation after P/I stimulation in Jurkat T cells. pIκBα levels were densitometrically quantified in relation to β-actin. Error bars, S.D.; n = 3 biological replicates were quantified; unpaired t test (two-tailed). *, p < 0.05; **, p < 0.01. F, upon P/I stimulation, IL-2 and TNFα cytokine secretion is attenuated after C25-140 treatment. Error bars, S.D.; n = 3 biological replicates; unpaired t test (two-tailed). *, p < 0.05; **, p < 0.01; D, DMSO.