Figure 4. Cleavage of chicken GSDMA leads to cell lysis.
(A.) Measurement of GFP fluorescence in HEK293T/17 cells co-transfected with GFP and either full length (FL) chicken GSDMA or its N-terminal (NT) pore-forming domain after 48 hours. Each point represents a separate transfection experiment. ****, p<0.0001 by student’s t-test. (B.) Cell lysis measured by LDH release in HEK293T/17 cells co-transfected with chicken caspase-1 and either WT or D244A chicken GSDMA. Gasdermin constructs in this study do not encode the 3xFLAG domain. Each point represents a separate transfection experiment. **, p<0.01; ****, p<0.0001 by 2 way ANOVA with Tukey’s multiple comparisons correction. (C.) Images of HeLa cells after transfection with the N-terminal domain of chicken GSDMA. Transfected cells display classic pyroptotic ballooning. SYTOX green staining of the nucleus indicates permeabilization of cell membranes.