FIGURE 3.

Silencing of HO-1 with siRNA abolished the effect of HO-1 induction in vivo. A, the distribution of FAM-siRNA in lung tissues isolated from DO11.10 mice. OCT-embedded lung sections were prepared 24 h after transfection (day −2) and 1 day after the last OVA challenge (6 days after the transfection (day 3)) and stained with DAPI. The FAM-expressing cells in lung tissues were visualized under a fluorescence microscope (original magnification ×400). B, histological analysis of lung tissues isolated from DO11.10 mice in control (CON), OVA, OVA + hemin + scrambled siRNA, and OVA + hemin + HO-1 siRNA groups. Paraffin-embedded lung sections were prepared 1 day after the transfection (day −2) and 1 day after the last OVA challenge (6 days after the transfection (day 3)) and stained with hematoxylin and eosin to observe inflammation (original magnification ×400). Each symbol in the graph represents an individual mouse (n = 6). C, Western blot analysis of HO-1 protein expression in lung tissues was extracted from the four groups. β-Actin was used as the loading control. Densitometry analysis was performed by normalizing to β-actin levels (* compared with the control (CON) group: *, p < 0.05; #, compared with the OVA group: #, p < 0.05). D, Western blot analysis of RORγt protein expression in lung tissues extracted from the four groups. β-Actin was used as the loading control. Densitometry analysis was performed by normalizing to β-actin levels (* compared with the control group: *, p < 0.05; # compared with the OVA group: #, p < 0.05). All results shown are representative of three independent experiments.