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. 2022 Dec 8;13(12):1029. doi: 10.1038/s41419-022-05476-3

Fig. 2. Lysosomal cathepsins modulate IL-1β production in response to flagellin independently of NLRP3 activation.

Fig. 2

Bone marrow-derived macrophages (BMDM) from C57BL/6j WT or Nlrp3−/− mice were pretreated with the cathepsin B inhibitor Ca-074Me (25 μM), primed with LPS (200 ng/ml, 3 h). Next, the cells were stimulated with silica (250 μg/ml, 6 h) or with ultrapure flagellin extracted from Salmonella typhimurium inserted into DOTAP (FliDot) (1 μg/ml, 6 h). A IL-1β secretion was evaluated in the culture supernatants by HTRF. The bars represent the average of three independent experiments performed in technical triplicates ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 (Three-way ANOVA) when compared to untreated cells. B Bone marrow-derived macrophages (BMDMs) from WT, Nlrp3−/−, Naip5−/−, Nlrc4−/−, and Aim2−/− mice were pretreated with the cathepsin B inhibitor Ca-074Me, primed with LPS and transfected with poly dA: dT (1 μg/ml, 16 h). Secretion of IL-1β was determined by electrochemiluminescence in the culture supernatant. The bars represent the mean ± SD of experimental triplicates. Data representative of two independent experiments.