Figure 2 | Cell Death & Disease

Figure 2

From: Cellular IAP proteins and LUBAC differentially regulate necrosome-associated RIP1 ubiquitination

Figure 2

RIP1 undergoes K63 and linear chain-linked polyubiquitination during necroptosis. (a) HT29 cells were treated with TNFα 20 ng/ml (T), BV6 2 μM (B) and zVAD (20 μM) for 3.5 hours. Cells were lysed in 6 M urea buffer and immunoprecipitated using linkage-specific anti-ubiquitin antibodies or control antibody. Immunoprecipitated proteins were detected using indicated antibodies. (b and c) MEF (b) or L929 (c) cells were not treated or treated with TNFα (100 ng/ml), BV6 (2 μM) and zVAD (20 μM) for 3 h (b) or with TNFα (10 ng/ml) and zVAD (20 μM) for 2 or 3 h (c). Cells were lysed and immunoprecipitated as in (a) and immunoprecipitated proteins were detected using indicated antibodies

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