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. 2023 Jan 10;13:1078526. doi: 10.3389/fimmu.2022.1078526

Figure 5.

Figure 5

Determination of the cleavage of duGSDME by ducaspase-3 and ducaspase-7 in eukaryotic cells. (A) HEK293 cells were transfected with vectors expressing ducaspase-3, ducaspase-3 without prodomain (npducaspase-3), or ducaspase-3 p10 and ducaspase-3 p17. The whole cell lysates of the transfected cells were incubated with Ac-DEVD-pNA. Eukaryotically expressed npducaspase-3 showed caspase-like enzymatic activity, while ducaspase-3 did not. **, p=0.01; ns, not significant. (B) Cleavage of duGSDME by ducaspase-3 in HEK293 cells. Cells were co-transfected vectors expressing npducaspase-3 and duGSDME. Western blot assay showed a 35 kDa protein could be recognized by anti-duGSDME-NT antibody besides the 55 kDa full-length duGSDME in the co-transfection group, but not in other groups. (C) Cleavage of duGSDME by ducaspase-7 in HEK293 cells. Cells were co-transfected vectors expressing ducaspase-7 and duGSDME. Western blot assay showed a 35 kDa protein could be recognized by both anti-duGSDME-NT antibody and anti-Flag antibody besides the 55 kDa full-length duGSDME in the co-transfection group, but not in other groups.