Fig 1: PLAU overexpression increased EGFR signaling in vitro. (A) Levels of PLAU, EGFR, p-EGFR, and ß-actin proteins in human bronchial epithelial (HBE) cells were measured by Western blotting analysis. Quantitative analysis of (B) PLAU, (C) EGFR, and (D) p-EGFR. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001.
Fig 2: PLAU overexpression reversed CSE-induced pyroptosis in vitro. (A) Levels of pro-caspase-1, cleaved-caspase-1, gasdermin D (GSDMD), GSDMD N-terminal domain, and ß-actin proteins in human bronchial epithelial (HBE) cells were measured by Western blotting analysis. (B) Levels of pro-IL-1ß and cleaved-IL-1ß in HBE cells were measured by Western blotting analysis. Quantitative analysis of (C) cleaved-caspase-1, (D) GSDMD N-terminal domain, and (E) cleaved- IL-1ß. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001.
Fig 3: PLAU knockdown reduced EGFR signaling in vitro. (A) Levels of PLAU, EGFR, p-EGFR, and ß-actin proteins in human bronchial epithelial (HBE) cells were measured by Western blotting analysis. Quantitative analysis of (B) PLAU, (C) EGFR, and (D) p-EGFR. mRNA levels of (E) PLAU, (F) EGFR. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001. shRNA vectors targeting PLAU.
Fig 4: GLIPR1 knockdown reduced PLAU/EGFR signaling in vitro. (A) Levels of GLIPR1, PLAU, EGFR, p-EGFR, and ß-actin proteins in human bronchial epithelial (HBE) cells were measured by Western blotting analysis. Quantitative analysis of (B) GLIPR1, (C) PLAU, (D) EGFR, and (E) p-EGFR. mRNA levels of (F) GLIPR1, (G) PLAU, (H) EGFR. (I) GLIPR1 was pull downed by Western blotting using anti- GLIPR1 antibody. (J) Immunoprecipitation assay showed that PLAU was pull downed by GLIPR1. (K) PLAU was pull downed by Western blotting using anti- PLAU antibody. (L) Immunoprecipitation assay showed that GLIPR1 was pull downed by PLAU. Error bars represent SD. *P < 0.05; **P < 0.01; ***P < 0.001. shRNA vectors targeting GLIPR1.
Fig 5: Glipr1 protected CS-induced airway damage through PLAU/EGFR signaling. (A) Levels of Glipr1, Plau, Egfr, p-Egfr, and ß-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.
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