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. 2017 Jul 17;36(17):2529–2543. doi: 10.15252/embj.201796476

Figure EV2. ZBP1 controls MCMV replication in NIH3T3 cells (related to Fig 2).

Figure EV2

  1. NIH3T3 cells reconstituted with wild‐type or mutant ZBP1 were infected as indicated. After 5 days, cells were fixed and stained with crystal violet.
  2. NIH3T3 cells reconstituted with wild‐type or mutant ZBP1 were infected as indicated using an MOI of 3. Control cells were treated with TZ. After 0, 8 or 16 h, cell lysates were subjected to Western blot analysis under non‐reducing conditions using the indicated antibodies. Short and long exposures of α‐MLKL Western blot are shown.
  3. CXCL10 was analysed by ELISA in supernatants from MEFs of the indicated genotypes infected with MCMV‐M45WT or MCMV‐M45mutRHIM (MOI = 3; 8 h). #not detected. The dotted line represents the lower limit of detection.
  4. HEK293T cells were transfected with 125 ng IFNβ firefly luciferase and 25 ng Renilla luciferase reporter plasmids, together with expression vectors for HA‐STING (500 ng), MDA5 (500 ng), RIPK3 (50 ng) or ZBP1‐3xFLAG (20, 100, 500 ng). Luciferase activity was measured after 24 h and the ratio of firefly and Renilla luciferase was set to 1 for control cells transfected with empty vector.
Data information: Data are representative of two or more independent experiments. Panels (C and D) show mean ± SD (n = 3). Source data are available online for this figure.