Fig. 2. Effect of TRAF3 deficiency on ISG transcription in T cells.

(A to C) WT and TRAF3^−/− HuT28 human T cells were stimulated with type I IFN (1000 U/ml) for the indicated times. (A) Induction of the expression of CXCL10, MX1, and STAT1 during type I IFN treatment, relative to that of a housekeeping gene [−ΔC_T = -(C_Tgene of interest- C_T housekeeping)], was determined by RT-qPCR analysis. (B) Top: Representative Western blotting analysis of STAT1 and MX1 proteins in the indicated cells. Bottom: Quantification of the normalized amounts of STAT1 and MX1 from seven biological replicates. (C) The amounts of CXCL10 protein secreted by the indicated cells were determined by ELISA. Each point represents an individual biological replicate. (D) Relative expression of Cxcl10, Ifitm3, and Ifit1 in type I IFN–stimulated control (WT) and Traf3^−/− mouse CD4⁺ T cells. Graphs include biological replicates from three (A and D) and seven (B and C) independent experiments with similar results. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by two-way ANOVA with Sidak’s multiple comparisons test. Error bars represent the SEM.