Figure 4.

LOX overexpression improves epithelial barrier integrity. (A) qRT-PCR for DSG1 and DSC1 in EPC2-hTERT organoids overexpressing GFP or LOX OE. GFP and LOX OE organoids were cultured with or without IL13 (10 μg/mL) from days 7 to 11 and then harvested on day 11 (n = 3). Data are representative of 3 independent experiments. (B) Schematic of ALI model. GFP and LOX OE EPC2-hTERT cells were cultured in low-calcium (0.09 mmol/L Ca²⁺) media for 3 days, followed by high-calcium media (1.8 mmol/L Ca²⁺) for 5 days, and then brought to ALI on day 8. ALI-cultured cells were stimulated with IL13 (10 μg/mL) from days 9 to 14. (C) TEER (Ω ∗ cm²) of the GFP and LOX OE EPC2-hTERT ALI cultures (n = 5). (D) Representative images of H&E staining, immunohistochemistry for TP63, and immunofluorescence staining for IVL (red), FLG (green), and DSG1 (green) of the GFP and LOX OE EPC2-hTERT ALI cultures. 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bar: 50 μm. Data are representative of 2 independent experiments and expressed as means ± SDs. (A) One-way analysis of variance and (C) 2-tailed Student t test were performed for statistical analyses. ∗P < .05, ∗∗P < .01. NT, nontreated.