Fig 2: GLIPR1 knockdown exacerbated CSE-induced pyroptosis in vitro. (A) Levels of pro-caspase-1, cleaved-caspase-1, gasdermin D (GSDMD), GSDMD N-terminal domain, and β-actin proteins in human bronchial epithelial (HBE) cells were measured by Western blotting analysis. (B) Levels of pro-IL-1β and cleaved-IL-1β in HBE cells were measured by Western blotting analysis. shRNA vectors targeting GLIPR1.

Fig 3: Glipr1 protected CS-induced airway damage through PLAU/EGFR signaling. (A) Levels of Glipr1, Plau, Egfr, p-Egfr, and β-actin proteins in the lung tissues of mice were measured by Western blotting analysis. Quantitative analysis of (B) Glipr1, (C) Plau, (D) Egfr, and (E) p-Egfr. (F) mRNA levels and (G) protein expression in bronchoalveolar lavage fluid (BALF) of Plau. (H) IHC. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001.

Fig 4: Glipr1 knockout aggravated caspase-1-dependent pyroptosis in CS-induced airway damage. (A) Levels of pro-caspase-1, cleaved-caspase-1, gasdermin D (GSDMD), GSDMD N-terminal domain, and β-actin proteins in the lung tissues of mice were measured by Western blotting analysis. Quantitative analysis of (B) cleaved-caspase-1 and (C) GSDMD N-terminal domain. (D) Levels of pro-IL-1β and cleaved- IL-1β in the lung tissues of mice were measured by Western blotting analysis. (E) Quantitative analysis of cleaved-IL-1β expression. (F) IL-1β concentration in bronchoalveolar lavage fluid (BALF) were measured by ELISA. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001.

Fig 5: Glipr1 knockout caused more severe inflammatory infiltration after cigarette smoke exposure. (A) Total protein levels, (B) total cell counts, (C) neutrophils, (D) macrophages, and (E) lymphocytes in bronchoalveolar lavage fluid (BALF) were assessed. IL-6 concentrations in (F) BALF and (G) plasma samples were measured by ELISA. (H) MMP-9 concentration in BALF were measured by ELISA. (I) Ratio of MMP-9/TIMP-1 in the BALF. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001.