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. 2016 Apr 7;1(4):e86355. doi: 10.1172/jci.insight.86355

Figure 7. Effect of calpain 14 overexpression on desmoglein 1 expression.

Figure 7

EPC2 cells transduced with empty vector (EV), calpain 14 overexpression vector (CAPN14), nonsilencing control (NSC), or CAPN14 gene-silencing (KD-1 and KD-2) vectors were grown at the air-liquid interface (ALI) and analyzed by (A) Western blot analysis for desmoglein 1 (DSG1). Quantitation of the 50-kDa DSG1 band intensity normalized to the full-length DSG1 band intensity is shown. (B) Immunofluorescence of DSG1 (green) in ALI culture; DAPI is shown in blue. (C) Western blot analysis of the CAPN14 gene-silencing effect on IL-13–mediated appearance of a DSG1 immunoreactive molecular species (50-kDa band) and quantitation of the 50-kDa DSG1 band intensity normalized to full-length DSG1 band intensity. (D) Immunofluorescence of DSG1 (green) in ALI-cultured EPC2 cells with and without IL-13 stimulation and CAPN14 gene silencing; DAPI is shown in blue. (E) ALI immunofluorescence intensity relationship of CAPN14 (green) and DSG1 (purple) and (F) colocalization of CAPN14 (green) and DSG1 (purple); DAPI is shown in blue. Data are representative of 3 independent experiments performed with replicates (n = 3–6). For A and C, data are expressed as the mean ± SEM; ***P < 0.001; statistical significance determined using a 2-tailed t test. Original magnification, ×20. Scale bar: 10 μm.