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. 2021 Jan 25;11:2158. doi: 10.1038/s41598-020-79607-1

Figure 2.

Figure 2

Pro-inflammatory markers and neutrophil recruitment in BAL-fluids (BALF) in PT + HS model. (A) Total protein in BALF. Kruskal–Wallis test with Dunn’s post-hoc analysis was performed. *** denotes p < 0.001 when PT + HS group is compared with its corresponding sham cohort. (B) PMN recruitment in BAL. Student’s t-test was performed to compare within PT + HS groups. PT + HS and sham animals were compared with Kruskal–Wallis test and *** denotes p < 0.001. (C) IL-6 content in BALF. Mann–Whitney rank sum test was performed to compare within the PT + HS groups. (D) TNF-α, (E) IL-1β and (F) IL-17 in BALF. For (E) and (F), Kruskal–Wallis test with Dunn’s post-hoc analysis was performed with * and ** denoting p < 0.05 and p < 0.01, respectively, when all PT + HS groups are compared to sham groups. For comparison between Ctrl-treated and NoxD21-treated PT + HS samples, Student’s t-test was performed. Animals per group for all measurements were n = 6–9. (G) CC16 in plasma. Two-way ANOVA with Student-Newman-Keul’s posthoc analysis was performed where * denotes p < 0.05 when PT + HS groups are compared to corresponding sham groups. Animals per group were n = 7–9. (H–K) Formalin-fixed, paraffin-embedded lung sections of Sham (Ctrl treated), Sham (NoxD21 treated), PT + HS (Ctrl treated) and PT + HS (NoxD21 treated) mice stained with H&E to characterize lung morphology. Intra-alveolar cell infiltration is indicated with a black arrow and RBCs are indicated with a blue arrow. A total of n = 8–12 lungs were stained for each group, and one representative image per group is presented. Images are at × 40 magnification. Bar = 50 µm. (L–O) Formalin-fixed, paraffin-embedded lung sections of Sham (Ctrl treated), Sham (NoxD21 treated), PT + HS (Ctrl treated) and PT + HS (NoxD21 treated) mice stained for myeloperoxidase (MPO) to detect neutrophil recruitment and activity within the lungs. Infiltrated granulocytes (marked with black arrow), alveolar epithelial cells and terminal bronchiolar epithelial cells are positively stained for MPO. A total of n = 4 lungs was stained for each group, and one representative image per group is presented. Images are at 40 × magnification. Bar = 100 µm.