FIGURE 5. Lanthanides do not block NLRP3 inflammasome activation or IL-1β release while Gsdmd deficiency also does not block NLRP3 inflammasome activation but does block IL-1β release.

(A) LPS-primed WT BMDM were stimulated with NG (10μM) for 30 min in the presence or absence of Gd³⁺ or La³⁺ (0.3, 1, 1.2, and 1.5mM). The ECM and soluble cell lysates were processed and analyzed on western blot for the presence of caspase-1 and IL-1β. The detergent insoluble fraction was DSS crosslinked and analyzed on western blot for the presence of oligomerized ASC. Western blot analysis of the soluble lysate for actin was also performed. These data are representative of results from 2 experiments. (B) LPS-primed WT BMDM were treated as in (A) in the presence of 5mM glycine. The ECM, soluble lysate, and detergent insoluble fraction were analyzed on western blot as described in (A). These data are representative of results from 2 separate experiments. (C) LPS-primed WT, Gsdmd^{−/− G1} (G1), and Gsdmd^{−/− G2} (G2) iBMDM were stimulated with NG for 30 min. The extracellular media (ECM) and soluble lysates were processed and analyzed on western blot for the presence of caspase-1 and IL-1β. Western blot analysis of the soluble lysate for Gsdmd and actin were also performed. The detergent insoluble fraction of the cell lysates were incubated with the chemical cross-linker disuccinimidyl suberate (DSS) and analyzed on western blot for the presence of ASC monomers, dimers, and oligomers. These data are representative of results from 2 experiments. *: non-specific band .