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. 2014 Dec 19;111(52):18673–18678. doi: 10.1073/pnas.1413687112

Fig. 5.

Fig. 5.

Preventive potential of tTbet-TMD in the alleviation of CIA. (A) Structure of tTbet-TMD. (B) Competitive inhibition of Tbet-dependent transcriptional activity by tTbet-TMD was analyzed in HEK293 cells cotransfected with the vectors expressing wild-type Tbet and luciferase driven by IFN-γ promoter. (C) The mouse primary naïve CD4+CD25−CD62Lhigh T cells were incubated with 1 or 4 μM of tTbet-TMD for 1 h, and then cells were stimulated with plate-bound anti-CD3 and soluble anti-CD28 antibodies for 48 h under TH1-polarizing conditions. The cells were stained for CD4 and IFN-γ, and then analyzed by FACS. (D and E) i.p. injection of MTX (35 mg/kg) and tTbet-TMD (2.5 mg/kg) were performed every other day for 4–8 wk from day 1 after primary immunization. (D) Inflammatory condition of the paws was observed before mice were killed. (E) Microscopic analysis of arthritis was assessed by paw thickness. Data are representative of at least three (B and C) independent experiments or eight mice per group (D and E). Error bars denote SEM. *P < 0.05, ***P < 0.001.