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. 2016 Jul 14;35(16):1766–1778. doi: 10.15252/embj.201694696

Figure 3. GSDMD Nterm targets liposomes after caspase‐1 cleavage.

Figure 3

  1. Human GSDMD at a concentration of 2 μM was incubated at room temperature with 5 nM caspase‐1. The protein was cleaved in a time‐dependent manner into two bands of 31 kDa (GSDMD Nterm) and 22 kDa (GSDMDCterm).
  2. Cross‐linking experiment of full‐length and cleaved GSDMD. GSDMD at a concentration of 2 μM was incubated at room temperature with 5 nM caspase‐1. After enzymatic cleavage, GSDMDNterm is highly cross‐linked by DSS, resulting in the gel‐impenetrating species highlighted by the arrow. GSDMDCterm is not cross‐linked.
  3. GSDMD at a concentration of 1 μM was incubated at room temperature with 5 nM caspase‐1 and liposomes composed of 4 mM DMPC or polar lipid extract derived from E. coli. After 2 h, the lipid fraction (L) was separated from the supernatant (S) by ultracentrifugation at 4°C for 1 h at 120,000 g.

Source data are available online for this figure.