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. 2021 May 11;9(5):535. doi: 10.3390/biomedicines9050535

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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LTB₄ receptors are necessary for NLRP3 stimulation and IL-1β production in HDM/LPS-induced neutrophilic airway inflammation. The control DMSO, U75302 (50 μg/kg) or LY255283 (10 mg/kg) were administered via i.p. injection 1 h before every challenge. Naïve designates control without any additions. (A) Levels of IL-1β in lung lysates were analyzed using ELISA. Data are shown as the mean ± SD (n = 57 per group). (B,C) The mouse lungs were homogenized, protein was isolated, and the levels of BLT1/2, NLRP3, caspase-1, and ASC complex were assessed by Western blotting. Data are representative of three independent experiments with similar results. (D) Immunofluorescence staining for NLRP3 (red, CY^TM3) in lung tissues. Nuclei were stained with DAPI (blue) (400×). The images are representative of three independent experiments with similar results. (E) The lungs were excised, fixed and stained with H&E and PAS. Peribronchial and perivascular lung inflammation and goblet cells were measured and scored (400×). Data are shown as the mean ± SD (n = 4–5 per group). (F) Levels of MPO in lung lysates were analyzed using ELISA. Data are shown as the mean ± SD (n = 5–7 per group). (G) Neutrophils, eosinophils, and total immune cells in BALF were obtained using Cytospin and stained with Diff-Quik. Data are shown as the mean ± SD (n = 4 per group). All experiments were performed in triplicate. ** p < 0.01 versus each control group.

LTB₄ receptors are necessary for NLRP3 stimulation and IL-1β production in HDM/LPS-induced neutrophilic airway inflammation. The control DMSO, U75302 (50 μg/kg) or LY255283 (10 mg/kg) were administered via i.p. injection 1 h before every challenge. Naïve designates control without any additions. (A) Levels of IL-1β in lung lysates were analyzed using ELISA. Data are shown as the mean ± SD (n = 57 per group). (B,C) The mouse lungs were homogenized, protein was isolated, and the levels of BLT1/2, NLRP3, caspase-1, and ASC complex were assessed by Western blotting. Data are representative of three independent experiments with similar results. (D) Immunofluorescence staining for NLRP3 (red, CY^TM3) in lung tissues. Nuclei were stained with DAPI (blue) (400×). The images are representative of three independent experiments with similar results. (E) The lungs were excised, fixed and stained with H&E and PAS. Peribronchial and perivascular lung inflammation and goblet cells were measured and scored (400×). Data are shown as the mean ± SD (n = 4–5 per group). (F) Levels of MPO in lung lysates were analyzed using ELISA. Data are shown as the mean ± SD (n = 5–7 per group). (G) Neutrophils, eosinophils, and total immune cells in BALF were obtained using Cytospin and stained with Diff-Quik. Data are shown as the mean ± SD (n = 4 per group). All experiments were performed in triplicate. ** p < 0.01 versus each control group.