Extended Data Fig. 4: Palmitoylation promotes GSDMD-NT-induced pyroptosis.
From: A palmitoylation–depalmitoylation relay spatiotemporally controls GSDMD activation in pyroptosis
a–d, HEK293T cells were transfected with the indicated constructs in the presence or absence of 2-BP (50 μM) for 24 hours. Cell death was measured by SytoxGreen positivity assay (a and c). The resulting supernatants were quantified for LDH release (b and d). Data are presented as mean ± s.d. of n = 5 (a and b), 4 (c), or 6 (d) independent wells of one representative experiment. Unpaired two-tailed Student’s t-test (a and b). One-way ANOVA post hoc Dunnett’s tests (c and d). e, f, HEK293T cells were transfected with the indicated constructs in the presence or absence of 2-BP (50 μM) for 24 hours. Cell lysates were then collected and subjected to Western blot analyses with the indicated antibodies. g, h, HEK293T cells were transfected with the indicated constructs in the presence or absence of 2-BP (50 μM) for 24 hours. Cell death was measured by SytoxGreen positivity assay (g). The resulting supernatants were quantified for LDH release (h). Data are presented as mean ± s.d. of n = 3 independent wells of one representative experiment (g and h). Unpaired two-tailed Student’s t-test (g and h). i, HEK293T cells were transfected with the indicated constructs in the presence or absence of 2-BP (50 μM) for 24 hours. Cell lysates were then collected and subjected to Western blot analyses with anti-GSDMD antibodies. j, k, HEK293T cells were transfected with the indicated constructs in the presence or absence of 2-BP (50 μM) for 24 hours. Cell death was measured by SytoxGreen positivity assay (j). The resulting supernatants were quantified for LDH release (k). Data are presented as mean ± s.d. of n = 3 independent wells of one representative experiment (j and k). One-way ANOVA post hoc Bonferroni’s tests (j and k).
