Fig. 3

E. histolytica-induced caspase-4 activation is independent of inflammasome components. a, b WT, NLRP3 CRISPR/Cas9 KO THP-1, and c, d THP-1 defASC 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 immunoblotted and immunoblot analysis was performed for caspase-4, caspase-1, and IL-1β in the supernatants (SN) and along with NLRP3 and ASC in lysates (LYS). Blots were reprobed for GAPDH. Quantifications of caspase-4 proteins were performed by densitometric analysis (b, d) 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 < 0.01). Bars represent ± SEM