Fig. 5: Cell lysis does not regulate caspase-4 cleavage and IL-18 production. | Communications Biology

Fig. 5: Cell lysis does not regulate caspase-4 cleavage and IL-18 production.

From: Gasdermin-D pores induce an inactivating caspase-4 cleavage that limits IL-18 production in the intestinal epithelium

Fig. 5

a–e HCT116 cells were electroporated then placed in media with 10 mM glycine to prevent cell lysis. In panels (f–i), HCT116 cells were silenced for NINJ1 expression using shRNA constructs, as indicated, before LPS electroporation. Cell lysis was measured by LDH cytotoxicity assay (a, g). GSDMD (b) and caspase-4 (e, i) cleavage was measured by western blots of whole cell lysates. IL-18 was measured by ELISA on supernatants (c), or on cells lysed into supernatant (d, h). The effect of NINJ1 shRNA constructs 1 and 5 on NINJ1 expression in HCT116 cells was determined by qRT-PCR (f). Green arrow indicates LPS dependent caspase-4 cleavage products. Representative of at least 3 experiments (except for (h) where n = 2), data displayed as mean ± S.E.M. *p < 0.05.

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