Fig. 7: Mincle is required for Fu-Hp-induced proinflammatory cytokine expression and NLRP3 inflammasome activation.

a The relative expression of indicated genes after Fu-Hp treatment in nonspecific (shNS) or CLEC4E knockdown (shCLEC4E) monocytes using qRT-PCR (n = 4 per group). Statistical significance was calculated with a one-way ANOVA with Bonferroni test and presented as means ± SD. arb. units, arbitrary unit. b Western blotting for mature IL-1β, mature caspase1, pro-IL-1β, and pro-caspase1 in human primary monocytes treated for 18 h with Fu-Hp. NM, Human Mincle neutralizing antibody; IgG, Mouse Control IgG2b. c, The concentrations of IL-1β in the supernatants of human primary monocytes with Fu-Hp treatment after indicated antibodies (NM or IgG) pretreatment measured by ELISA (n = 4 per group). Statistical significance was calculated with a two-sided unpaired t-test with Mann-Whitney test and presented as means ± SD. d, The relative expression of indicated genes after Fu-Hp treatment in mouse BMDM from Clec4e^WT or Clec4e^KO using qRT-PCR (n = 4 per group). Statistical significance was calculated with a two-sided unpaired t-test with Mann-Whitney test and presented as means ± SD. arb. units, arbitrary unit. ns, not significant. e Western blotting for mature IL-1β, mature caspase1, pro- IL-1β, IL-6, and pro-caspase1 in mouse BMDM from Clec4e^WT or Clec4e^KO treated for 18 h with Fu-Hp. f The concentrations of IL-1β, IL-6, and TNF in the supernatants of Fu-Hp-treated Clec4e^WT or Clec4e^KO BMDM (100 μg/mL for 18 h) measured by ELISA (n = 4 per group). ns, not significant. Statistical significance was calculated with a two-sided unpaired t-test with Mann-Whitney test and presented as means ± SEM. g Firefly luciferase reporter vector (NFAT-RE) were stimulated with HC-Hp or Fu-Hp, and reporter assay was performed in HeLa cells transfected with a HA-Mincle expression vector (left; n = 4 per group). HA-Mincle transfection was confirmed by WB using an HA antibody (right). Statistical significance was calculated with a two-sided Kruskal-Wallis test and presented as means ± SD. RLU/s, Relative Light Units per second.