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

Figure EV3. Validation of ZBP1‐Zα1α2mut knock‐in (related to Fig 3).

Figure EV3

  1. Targeting strategy. See Materials and Methods for further details.
  2. DNA fragments encompassing exon 2 or 3 were PCR‐amplified from genomic DNA from primary MEFs of the indicated genotypes and were then sequenced.
  3. Primary MEFs of the indicated genotypes were treated with 100 U/ml IFN‐A/D for 16 h. RNA was extracted, and Zbp1 expression was analysed by RT qPCR.
  4. Cell death was monitored upon treatment with TZ using an in‐incubator imaging platform (Incucyte) and the dye Sytox Green that stains dead cells.
  5. Non‐reducing SDS–PAGE and Western blot for MLKL from samples from Fig 3D.
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.