Figure 4.

Pretreatment with rLCN2 promotes lung oxidative stress and iron deposition in LPS-treated WT and LCN2 KO mice. (A) Western blotting and quantification of lung HO-1, 4-HNE, and SOD-2 protein levels in lung tissues. β-actin served as a loading control. (n = 5-7). (B) Representative images of Perls Prussian blue iron staining in lung sections and BALF slides. Arrows indicate iron-stained alveolar macrophages. Scale bar, 50 μm. (n = 4 mice). (C) In lung sections and BALF slides (B), iron-positive alveolar macrophages were counted and analyzed in four fields (100x100 μm²) for each slide (n = 6). (D) Quantification of total lung iron levels using an iron assay kit. (n = 6 mice). Differences between two groups were evaluated using unpaired Student's t-tests. *P < 0.05 versus LPS-treated WT. †P < 0.05 versus rLCN2+LPS-treated WT. All data are presented as mean ± SEM.