Fig. 5

E. histolytica-induced IL-1β secretion is dependent on caspase-4 and -1 activation. a WT, CASP4 CRISPR/Cas9 KO and CASP1 CRISPR/Cas9 KO THP-1 macrophages were incubated with Eh for 60 min or MSU (300 μg/mL for 6 h) and cell supernatant was quantified by human focused 13-plex cytokine/chemokine arrays. b Cell supernatant was added to HEK-Blue™ reporter cells to detect bioactive IL-1β using the SEAP assay. c Cell supernatant was used to measure cell death by LDH release and is shown as a percentage of LDH release from cells without stimulation (control). LN is abbreviated for LPS (50 ng/mL) and nigericin (10 μM) stimulation for 30 min. d WT, CASP4 CRISPR/Cas9 KO and CASP1 CRISPR/Cas9 KO THP-1 macrophages were incubated with LPS (50 ng/mL) and nigericin (10 μM) stimulation for 60 min. Equal amounts of lysates were loaded onto SDS-PAGE gel and immunoblot analysis was performed for caspase-4, caspase-1, IL-1β in both the supernatants (SN) and lysates (LYS). Blots were reprobed for GAPDH. e Cell supernatant was added to HEK-Blue™ reporter cells to detect bioactive IL-1β using the SEAP assay. Data are representative of three experiments and statistical significance was calculated with ANOVA and Bonferroni’s post-hoc test (***p < 0.001). Bars represent ± SEM