Total cell lysates of cultured cells were prepared using M-PER (Thermo Fisher Scientific, Runcorn, Cheshire, United Kingdom) Primary antibodies against pRIPK3, RIPK3, pMLKL, and MLKL from a Mouse Reactive Necroptosis Antibody Sampler Kit (Cell Signaling Technology) were used. After incubation with secondary antibodies, immunocomplexes were detected using a WB7108; Thermo Fisher Scientific. β-actin was used as an internal control.