Fig. 4

E. histolytica-induced caspase-1 activation is dependent on caspase-4 and caspase-4 activation is independent of caspase-1. a–c WT, CASP4 CRISPR/Cas9 KO and d, e CASP1 CRISPR/Cas9 KO THP-1 macrophages were incubated with Eh (1:20) at increasing time points. Cell supernatant was TCA precipitated and cells were washed and lysed. 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. Quantifications of caspase-1 and IL-1β (b and c) and caspase-4 (e) were performed by densitometric analysis from three independent experiments, and the negative (cells only) acted as an internal control. Data are representative of three experiments and statistical significance was calculated with Student’s t-test between KO and WT (*p < 0.05). Bars represent ± SEM