Fig. 6

VLP-induced release of inflammatory molecules is related to NLRP3 inflammasome activation. Macrophages were treated with VLPs (20 µg/ml) and mAbs (7.5 µg/ml) for 24 h. siRNA was used to downregulate NLRP3 gene expression, negative (neg.) siRNA was used as a control. WB of NLRP3 protein expression: A representative image, B quantification (N = 5), to calculate relative NLRP3 expression NLRP3 protein signal was normalized to β-actin signal and the calculated value was divided by control value in the absence of siRNA (No siRNR equal to 1.0 of every independent experiment). C, E IL-1β (N = 10 and 8) and D, F TNF-α (N = 9 and 8), (H) CXCL1 (N = 5), (I) CXCL2 (N = 6), (J) CXCL9 (N = 10), (K) CCL8 (N = 5) secretion determined by ELISA. Inhibitor MCC950 to block NLRP3 inflammasome activation was used at a concentration of 1 μM and added 30 min before the treatment (E). (G) IL-1β (N = 4–9) secretion induced by different IC, mAb clone refers to IC formed by VLPs and the respective mAb. Data are represented using bar graphs with dots showing the number of independent experiments, numbers above the bars are mean values, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, one-way ANOVA followed by Tukey’s multiple comparison test with option of matched measures across one N