Figure 3. MLKL ubiquitylation accumulates in the crude membrane fraction and can be digested by USP21 located on biological membranes.

1. WT MDFs were treated with TSI for indicated time. Cells were fractionated into cytosol (C) and crude membrane (M), followed by UBA pulldown. All fractions were analysed by Western blot and probed with antibodies as indicated. Representative of three independent experiments.
2. WT USP21‐CaaX and USP21^C221R‐CaaX (CR) were inducibly expressed in WT MDFs by doxycycline for 5 h. Ubiquitylated proteins were enriched followed by TSI stimulation and cellular fractionation. All fractions were analysed by Western blot and probed with antibodies as indicated. Representative of three independent experiments.

Data information: Wild‐type (WT), whole cell lysate (WCL). TSI is used as a necroptotic stimulus.

Source data are available online for this figure.