Figure 5.
Plasma membrane translocation of trimerized MLKL in response to necroptosis induction. (a) Control shRNA or MLKL shRNA HT29 cells (clone 1-3-2) were treated with TSZ. The total, cytosolic and crude membrane fractions were resolved either on reducing or non-reducing gel and analysed by immunoblotting as indicated. T, total cell lysate. C, cytosolic fraction. M, membrane fraction. (b) HT29 cells were treated with TSZ as indicated. Total cellular lysate and the biotinylated, cell surface fraction were resolved on reducing gel and analysed by immunoblotting as indicated. (c) HEK293 cells were transfected as indicated. Total cellular lysate and the biotinylated, cell surface fraction were resolved on non-reducing gel and analysed by immunoblot as indicated. Δ indicates phosphorylated RIP3. Data shown are representative of two independent experiments. Uncropped images of western blots are shown in Supplementary Fig. 8.