Figure 8.

Role of p53 and PAI-1 in CS-induced MH and cilia dysfunction in mice. A: AECs isolated from nL were transduced with Lv-p53 shRNA or control nonspecific shRNA. These cells were treated with or without CSE. AEC lysates were immunoblotted for MUC5AC, HDAC6, FOXA3, acetylated tubulin (Ac-Tub), CAV1, p53, PAI-1, and LC3BII. Same samples were analyzed for β-actin to assess equal loading. B: AECs from nL transduced with Ad-Ev or Ad-p53 were left untreated or treated with CSP7. Lysates of Ad-Ev, Ad-p53, and Ad-p53+CSP7 treated AECs were immunoblotted for listed proteins. C: AECs from nL transduced with Lv-PAI-1 shRNA or control nonspecific shRNA were later treated with or without CSE. The lysates were tested for above proteins by Western blotting. D: AECs transduced with Ad-Ev or Ad-PAI, and Ad-PAI-1-exposed AECs were later treated with or without CSP7. Lysates were immunoblotted for listed proteins. E: AECs isolated from ambient Air kept or CS-exposed (CS) WT, and p53- and PAI-1-deficient mice were immunoblotted for MUC5AC, HDAC6, Ac-Tub, CAV1, LC3BII, and β-actin. F: lung sections of WT, and p53- and PAI-1-deficient mice treated as in E were subjected to IHC staining for MUC5AC (scale bar 100 µM). AECs, airway epithelial cells; CAV1, caveolin‐1; CS, cigarette smoke; CSE, cigarette smoke extract; CSP7, caveolin-1 scaffolding domain peptide; HDAC6, histone deacetylase 6; MH, mucus hypersecretion; MUCAC, mucin 5AC; nL, “normal” lung; PAI-1, plasminogen activator inhibitor-1; WT, wild type.