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. 2019 Aug 7;38(18):e102075. doi: 10.15252/embj.2019102075

Figure 4. TRAF3IP3 mediates TRAF3 recruitment to MAVS upon virus infection.

Figure 4

  1. pcDNA‐FLAG‐TRADD and pcDNA‐FLAG‐TRAF3IP3 were transfected into HEK293T cells at increasing amount, respectively. ISG54 and IL6 induction were measured 36 h post‐transfection by qPCR. Expression levels are shown in Fig EV4B. All data are presented as the mean values based on three independent experiments, and error bars indicate s.d. P values were determined by unpaired two‐tailed Student's t‐test. *P < 0.05, **P < 0.01, and ***P < 0.001. NS indicates no statistically significant difference.
  2. HEK293T cells were transfected with pcDNA3‐HA‐TRAF3IP3, in combination with pcDNA3‐FLAG‐ RIG‐I, MAVS, TBK1, IRF3, TRAF2, TRAF3, TRAF5, or TRAF6 as indicated. Thirty‐six hours after transfection, cells were collected and anti‐HA immunoprecipitations were performed. IP products were subjected to immunoblotting. Protein expression levels are shown in Fig EV4C.
  3. sh‐RNA targeting TRAF3IP3 (sh‐TRAF3IP3) was transduced into Mavs −/− HEK293T cells. Twenty‐four hours after transduction, cells were treated with puromycin (2 μg/ml) for 48 h and then transfected with pcDNA3‐FLAG‐MAVS‐(Region III only). Twenty‐four hours after transfection, cells were infected with or without VSV for 12 h. Cells were then collected and subjected to immunoprecipitation assay and immunoblotting.
  4. Wild‐type and Traf3ip3 −/− HEK293T cells were transfected with pcDNA3‐FLAG‐MAVS‐(Region III only). Twenty‐four hours post‐transfection, cells were infected with or without VSV. Twelve hours post‐infection, cells were collected and subjected to immunoprecipitation assay and immunoblotting.

Source data are available online for this figure.