Figure 2.

Proinflammatory Activity of FlgE in Mice (A–D) or Lung Organoid (E). (A–D) C57BL/6 mice were intranasally administered 120 μg recombinant FlgE, FlgEM, or LPS in a 150-μL volume. Twenty-four hours later, the animals were sacrificed by bleeding under anesthesia, and bronchoalveolar lavage was performed before the lungs were cut into halves that were subjected to either fixation for hematoxylin/eosin staining (A) or to RNA extraction for RT-qPCR (C). Quantification of chemokines in the lavage was by ELISA (D), and myeloperoxidase (MPO) levels in the serum were measured using an MPO determination kit (B). Data are representative of two independent experiments that showed similar results. N = 6 mice for each treatment group. (E) For lung organoid stimulation, lung slices were treated with various reagents for 24 hours and then subjected for chemokine mRNA measurement with RT-qPCR. Data are representative of three independent experiments. Three mice were used for making mixture of lung organoid cultures. Two samples were analyzed for each group. *P < 0.05, **P < 0.01, ***P < 0.001 vs. Control/PBS. ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs. FlgE.