Tumor Necrosis Factor (TNF) Receptor Expression Determines Keratinocyte Fate upon Stimulation with TNF-Like Weak Inducer of Apoptosis

Figure 1

Detection of Fn14/TRAF2/cIAP1/TNFR complex in keratinocytes. Human primary keratinocytes were cultured in vitro, followed by extraction of protein lysates. Some cells were pretreated with TWEAK (100 ng/ml, 24 h). Using immunoprecipitation and Western blotting, we detected associations between the different potential binding partners. Proteins were precipitated by anti-TRAF2 (a), TNFR1 (b), cIAP1 (c), or Fn14 (d) IgG and then detected by other antibodies. Similarly, proteins were extracted from TNFR2-overexpressing keratinocytes and precipitated with anti-TRAF2 (e), TNFR2 (f), cIAP1 (g), or Fn14 (h) IgG. Input refers to the cell lysate that was aspirated before the addition of immunoprecipitation IgG. In contrast, control IgG refers to an antibody that is not related to the target protein. Data were obtained from five independent experiments, and none of the blots was stripped and reprobed. Representative images are shown.

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