Mouse rip3

TNF-α recombinant protein was generated as previously described.^{1 (link)} z-VAD was from Bachem (Babendorf, Switzerland). Necrostatin-1 was purchased from Alexis Biochemicals (San Diego, CA, USA). The following antibodies were used for western blot analysis: RIP1 (BD Biosciences, NJ, USA, 610458), phospho-MLKL (Abcam, Cambridge, UK, 187091), caspase-8 (Cell Signaling, Danfoss, MA, USA, 9746), CYLD (Cell Signaling, 437700), mouse RIP3 (Prosci (San Diego, CA, USA), 2283), Sp1 (Santa Cruz, CA, USA, 14027), UHRF1 (BD Biosciences, 612264), Dnmt1 (Abcam, 13537), Flag (Sigma, St. Louis, MO, USA, A8592) and β-actin (Sigma, A2066). The following antibodies were used for flow cytometry analysis: CD11b (Biolegend, San Diego, CA, USA, 101212), CD45 (BD, 563891), GR-1 (BD Biosciences RB6-8C5), F4/80 (Biolegend, 123113).

Dulbecco's modified Eagle's medium (DMEM) was from Thermo. Penicillin/streptomycin, L-Glutamine, and fetal bovine serum (FBS) were from GIBCO. BHA, NAC, phosphate buffered saline (PBS), and Lanthanum(III) chloride heptahydrate (LaCl₃·7H₂O) were from Sigma. Recombinant TNF-α was purified as described previously [11 (link)]. z-VAD was from Bachem. Necrostatin-1 was from Alexis Biochemicals. Propidium Iodide was from Biouniquer. The following antibodies were used for western blotting: mouse RIP3 (Prosci, 2283), RIP1 (BD Biosciences, 610459), mouse CYLD (Cell Signaling, 437700), caspase-3 (Cell Signaling, 9662), and β-actin (Sigma).