Figure 6. Effect of calpain 14 gene silencing on epithelial cell responses to IL-13.
EPC2 cells transduced with either a nonsilencing control (NSC) or calpain 14 (CAPN14) gene silencing (KD-1 and KD-2) vector were grown at the air-liquid interface (ALI) with and without IL-13 and analyzed by (A) quantitative PCR of CAPN14 mRNA relative to GAPDH mRNA and normalized to NSC without IL-13 (n = 3). (B) Western blot analysis of CAPN14 protein in ALI culture (n = 3) with HSP-90 as a loading control. (C) H&E staining of ALI-cultured EPC2 cells. (D) Quantification of the percentage of total area of dilated intercellular spaces. (E) Quantification of nuclei of basal cells in contact with basolateral edge of ALI epithelium. (F) Transepithelial resistance (Rt, n = 3) and (G) FITC-dextran flux (n = 3) of ALI system. Data are representative of 3 independent experiments. Scale bar: 10 μm. For A and D–G, data are represented as the mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; statistical significance determined using a 2-tailed t test.