Figure 2.

Effects of HMGB1 on tight junction molecules of 2D culture of HLE cells with 10% FBS and characterization of 2.5D Matrigel culture of HLE cells. (a) Western blotting for CLDN-1, −2, −4, −7, LSR, TRIC and actin in 2D culture of HLE cells pretreated with 10 μM EW-7197 2 h before treatment with 100 ng/ml HMGB1 for 24 h. The corresponding expression levels of A are shown as bar graphs. control vs *p < .05, TGF-β vs #p < .05. (b) Immunocytochemistry for OCLN and LSR in 2D culture of HLE cells (with 10% FBS) pretreated with 10 μM EW-7197 2 h before treatment with 100 ng/ml HMGB1 for 24 h. Scale bar, 20 μm. (c, d) Phase-contrast images and immunocytochemistry for OCLN and LSR and epithelial permeability assay using FITC-4kDa dextran (FD-4) in 2.5D Matrigel culture of HLE cells. (c) Bronchial-like spheroid cells. (d) alveolar-like spheroid cells. Scale bar, 20 μm