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. 2018 Jun 6;37(14):e97072. doi: 10.15252/embj.201797072

Figure 4. TRAF2 interacts directly with caspase‐2 to induce activation.

Figure 4

  1. A diagram of the caspase‐2 protein showing the major domains and the location of a putative TRAF‐interacting motif (TIM).
  2. Casp2(C320A)‐Flag plasmids (wild type or TIM mutant) were expressed in HeLa cells for 24 h followed by anti‐Flag IP and IB.
  3. Stably transfected, catalytically active caspase‐2 (wild type or TIM mutant) was induced in HeLa Tet‐On cells with 0.1 μg/ml doxycycline for 24 h, followed by propidium iodide (PI) staining and flow cytometry. n = 3 independent experiments (means + s.e.m.). *P < 0.05 by unpaired two‐tailed t‐test (top). Casp2‐Flag expression was confirmed by IB (bottom).
  4. Indicated amounts of Casp2(C320A) BiFC constructs were transiently transfected, and cells were cultured for 24 h followed by 20 μM cisplatin treatment in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h, and then, BiFC signal was detected by flow cytometry. n = 3 independent experiments (means + s.e.m.). *P < 0.05 by unpaired two‐tailed t‐test.

Source data are available online for this figure.