Fig. 5

Curdlan increases but wedelolactone downregulates caspase-11 activation and GSDMD cleavage in HDM-induced MH-S cells. MH-S cells were stimulated with PBS (50 μg/ml), HDM (50 μg/ml), HDM plus Laminarin (Lam) (1 μg/ml), HDM plus Curdlan (Cur) (200 μg/ml), or giving wedelolactone (Wed) (30 μM) after HDM/Curdlan for 24 h. A Total proteins were extracted and subjected, western blot was used to detect the expression of pro caspase-11 and cleavage caspase-11 in each stimulation group. B Densitometric quantification of immunoblots in (A). Levels of cleavage caspase-11 were normalized to GAPDH. C Expression of GSDMD-FL, GSDMD-N and NLRP3 in each stimulation group by western blot. D Densitometric quantification of immunoblots in (C). Levels of GSDMD-N were normalized to β-tubulin. E Densitometric quantification of immunoblots in (C). Levels of NLRP3 were normalized to β-tubulin. F Representative images of caspase-11 expression in each stimulation group detected by immune fluorescence microscopy. G, H The expression of IL-1α and IL-1β mRNA in the cells of each stimulation group by qRT-PCR. MH-S cells were stimulated with PBS (50 μg/ml), HDM (50 μg/ml), HDM plus Curdlan (200 μg/ml), or giving MCC950(10 μM) after HDM/Curdlan for 24 h, cleavage caspase-11 (I) and GSDMD-N (J) in each stimulation group by western blot. (*P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001, one way ANOVA, error bars represent mean ± SEM)