Figure 1

Localization of Nrf2 in small airway epithelium of healthy nonsmokers and healthy smokers. Cytospin preparations of small airway epithelium were assessed by immunohistochemical analysis and Western analysis. (A–F) Immunohistochemistry. (A, B) Healthy nonsmokers, + Nrf2 antibody; (C) healthy nonsmoker, + Nrf2 antibody + blocking peptide; (D–E) healthy smokers, + Nrf2 antibody; (F) healthy smoker, + Nrf2 antibody + blocking peptide. For (D–E), arrows indicate nucleoli positive for Nrf2 staining. For (A–F), bar = 10 µm. (G–J) Western analyses. For the Western analyses, small airway epithelial cells of four healthy nonsmokers and four healthy smokers provided nuclear and cytoplasmic extracts, which were normalized to equal protein concentration. (G) Western analysis of Nrf2 in cytoplasmic extracts from small airway epithelial cells of healthy nonsmoker and smoker (upper panel) and β-tubulin as an equal loading control (lower panel). Lane 1–4, examples of nonsmokers; lanes 5–8, examples of smokers. (H) Western analysis of Nrf2 in nuclear extract from small airway epithelium from healthy nonsmoker and healthy smoker (upper panel) and lamin B as loading control (lower panel). Lanes 9–12, examples of nonsmokers; lanes 13–16, examples of smokers. (I) Quantitative analyses of band densities for Nrf2 in cytoplasm (normalized to β-tubulin). (J) Quantitative analysis of band densities for Nrf2 in nucleus (normalized to lamin B). Data shown are mean ± standard error.