Fig. 5: Fu-Hp induces inflammatory responses and NLRP3 inflammasome activation in monocytes and macrophages.
From: Fucosylated haptoglobin promotes inflammation via Mincle in sepsis: an observational study
a The relative expression of IL1B, IL6, TNF, IL10, IL12B, CCL2, and CXCL2 in monocytes from a HC after treatment with HC-Hp or Fu-Hp (20 μg/mL for 6 h) using qRT-PCR (n = 3 per each group). Statistical significance was calculated with a two-tailed unpaired t-test and presented as means ± SD. arb. units, arbitrary unit. ns, not significant. b, The relative expression of Il1b, Il6, and Tnf in mouse BMDMs after treatment with HC-Hp or Fu-Hp (100 μg/mL for 6 h) using qRT-PCR (n = 4 per each group). Statistical significance was calculated with an unpaired t-test with Mann-Whitney test and presented as means ± SEM. arb. units, arbitrary unit. c,d The concentrations of IL-1β, IL-6, and TNF in the supernatants of HC-Hp or Fu-Hp-treated cells measured by ELISA (n = 4 per group). Statistical significance was calculated with an unpaired t-test with Mann-Whitney test and presented as means ± SEM. e, The relative expression of Il1b, Il6, Tnf, Il10, and Ccl2 in RAW264.7 cells after treatment with HC-Hp or Fu-Hp (20 μg/mL for 6 h) using qRT-PCR (n = 2 per each group). f,g Western blotting for phosphorylated NF-κB (p-p65), phosphorylated ERK (p-ERK), phosphorylated JNK (p-JNK), phosphorylated Akt (p-Akt), and phosphorylated p38 (p-p38) in human primary monocytes (f) or mouse BMDMs (g). h 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 after LPS priming. i The concentrations of IL-1β and TNF in the supernatants of human primary monocytes with Fu-Hp treatment after shNS or shNLRP3 virus knockdown 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. j The concentrations of IL-1β in the supernatants of human primary monocytes with Fu-Hp treatment after mitoTEMPO pre-treatment or not measured by ELISA (n = 4 per group). Statistical significance was calculated with a one-way ANOVA with Bonferroni test and presented as means ± SD (k) The concentrations of IL-1β in the supernatants of human primary monocytes with Fu-Hp treatment after potassium chloride (KCl) pre-treatment or not measured by ELISA (n = 4 per group). Statistical significance was calculated with a one-way ANOVA with Bonferroni test and presented as means ± SD.
