Figure 2
CS exposure induces MUC5AC, SPDEF, and Bcl-2 levels in human AEC monolayers. Primary human AECs grown in submerged cultured conditions were treated with 0, 1, 10, and 100 µg/ml of CSE for 24 h and the mRNA levels of MUC5AC (A) SPDEF (B) and Bcl-2 (C) were analyzed by qRT-PCR. (D) Immunoblot analyses of Bcl-2 expression following CSE exposure. Differentiated human AECs were treated with 10 µg/ml CSE and cell lysates were analyzed at 0, 24 and 48 h post CSE exposure by western blot analysis for Bcl-2 protein levels with β-actin levels detected as the loading controls. (E) Relative quantities of Bcl-2 protein as determined by densitometric analysis and normalized to β-actin levels. (F) Representative micrographs showing MUC5AC (green) and Bcl-2 (red) immunopositivity in HAECs treated with CSE or left non-treated (NT). Human AECs were treated with CSE (10 µg/ml) for 48 h and immunostained for Bcl-2 and MUC5AC, and nuclei were stained with DAPI (Scale – 5 µ). (G) Quantification of human AECs immunopositive for MUC5AC and Bcl-2 following CSE exposure. The percentage of human AECs immunopositive for Bcl-2 (Bcl-2+) and MUC5AC (MUC5AC+) were calculated. Data shown as mean ± SEM (n ≥ 3); *p < 0.05; **p < 0.01; ***p < 0.001.
