Figure 6.

Global DNA demethylating agent AZA restores lung pulmonary remodeling in both mouse and human lungs. (A) Global DNA methylation in WT C57BL/6J-derived mTECs after acute exposure to CS (two cigarettes or humidified air in a day) (n = 7 from two independent experiments). (B) Cytotoxicity in mouse precision-cut lung slices (PCLSs) from WT C57BL/6J with acute exposure to CS (six cigarettes or humidified air in a day) by LDH level (seven to nine slices from three independent experiments). (C and D) Immunofluorescence of (C) 5-methylcytosine (5mC; red) and nucleus (blue) and of (D) E-cadherin (green), 5hmC (red), and nucleus (blue) of PCLSs derived from WT mice (n = 3 from three independent experiments). Negative controls against rabbit IgG (Rb IgG) and mouse IgG (Ms IgG) are indicated at the bottom. (E and F) Immunofluorescence of (E) 5mC (red) and nucleus (blue) and of (F) E-cadherin (green), 5hmC (red), and nucleus (blue) in human PCLSs derived from patients with COPD (n = 3 from 2 donors). Relative fluorescent intensity is indicated on the right in (E) and at the bottom in (F). The immunofluorescence images were taken at a 63× oil objective. Scale bars, 20 μm. Error bars represent SEM. Statistics were determined by using (A, E, and F) the Mann–Whitney test and (B) one-way ANOVA with Sidak’s multiple comparison tests, with P < 0.05 considered statistically significant. LDH = lactate dehydrogenase.