Fig. 6

Overexpression of caspase-4 rescues E. histolytica-induced IL-1β secretion in CASP4-deficient macrophages. a CASP4 CRISPR/Cas9 KO THP-1 macrophages were nucleofected with caspase-4 expression plasmid (abbreviated pC-4) and left to rest for 24 h before incubating with Eh (1:20) for 60 min. b Cell supernatant from stimulated macrophages was also added to HEK-Blue™ reporter cells to detect bioactive IL-1β using the SEAP assay. Cell supernatant was TCA precipitated and cells were washed and lysed. Equal amounts of protein were loaded onto SDS-PAGE gel and immunoblot analysis was performed for caspase-4, caspase-1, and IL-1β in both the supernatants (SN) and lysates (LYS), and blots were reprobed for GAPDH. Data are representative of three experiments and statistical significance was calculated with Student’s t-test (**p < 0.01, ***p < 0.001). Bars represent ± SEM