Figure 2.

LCN2 deletion attenuates lung oxidative stress and iron accumulation in LPS-treated mice. (A) Western blot and quantification of HO-1 and SOD-2 protein levels. β-actin served as a loading control. (n = 4-7). (B) Representative double immunofluorescence images of CD11b and 4-HNE in lung sections. Nuclei were stained with DAPI. Scale bar, 50 μm. (C) Co-localized CD11b and 4-HNE-positive cells were counted and analyzed in three fields (100x100 μm²) for each slide (n = 6). (D) Representative images of Perls Prussian blue staining in lung sections and BALF slides. Arrows indicate iron-stained alveolar macrophages. Scale bar, 50 μm. (E) In lung tissues and BALF slides (D), iron-stained macrophages were counted and analyzed in four fields (100x100 μm²) for each slide (n = 6). (F) Representative triple immunofluorescence images of F4/80, LCN2, and ferritin in lung sections. Scale bar, 50 μm. (G) Co-localized F4/80, LCN2, and ferritin-positive cells were counted and analyzed in three fields (100x100 μm²) for each slide (n = 6). Differences between four groups were evaluated using two-way ANOVA followed by Tukey's multiple comparisons test. *P < 0.05 versus WT saline. †P < 0.05 versus WT LPS. All data are presented as mean ± SEM.