Fig. 5: IAMP29 activates the NLRP3 inflammasome via an interaction with PKM2.

a Heatmap showing the top protein hits identified by the LC‒MS analysis in PMA-differentiated THP-1 cells with FLAG or FLAG-IAMP29. b Volcano plot depicting the significant increase in PKM gene expression in A30L-treated human primary PBMCs. c Venn diagram illustrating the overlap of PKM2 in the three indicated groups. d, e Validation using PKM1 and PKM2 antibodies to detect immunoprecipitates from FLAG- or FLAG-IAMP29-treated cells (PMA-differentiated THP-1 and RAW 264.7 cells). f IL-1β concentrations in the supernatants of the indicated cells treated with IAMP29 (25 μg/mL for 18 h) after pretreatment with shikonin (5 μM for 2 h), as measured by ELISA. g Western blots showing the levels of mature IL-1β and mature caspase-1 in culture supernatants and pro-IL-1β, caspase-1, and β-actin in lysates of IAMP29-treated monocytes cultured in the absence or presence of shikonin (5 μM for 2 h). h IL-1β concentrations in the supernatants of IAMP29-treated monocytes (25 μg/mL for 18 h) with PKM2 knockdown, as measured by ELISA. IL-1β concentrations in the supernatants of shikonin-treated (i) or PKM2-knockdown monocytes (j) treated with the indicated NLRP3 inflammasome activators were measured by ELISA. k Western blots showing the levels of mature IL-1β and mature caspase-1 in culture supernatants and PKM2, pro-IL-1β, pro-caspase-1, and β-actin in the lysates of IAMP29-treated monocytes after PKM2 knockdown. Statistical analysis was conducted with an unpaired t-test with the Mann‒Whitney test or one-way ANOVA with Tukey’s multiple comparison test, and the results are presented as the means ± SDs. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.