Fig. 2.
Effect of chronic exposure to HDM in a KrasG12D-driven LC model. A. KrasG12D mice were treated i.n with VEH (n = 7), HDM (n = 8), or HI-HDM (n = 7) as indicated in this schematic overview of the study design. B Relative body weight of the mice. C Survival curves of mice treated as in A with VEH (n = 13), HDM (n = 19), or HI-HDM (n = 8). Death events occurring within the first 4 weeks after the beginning of the treatments were excluded. Values are expressed as a percentage of survival. D BALF total cell counts. E Representative pictures of H&E-stained lung sections. The lower panels are the same images as the above panels after tumor area quantification using QuPath software. QuPath-pseudocolored areas are represented as tumors (blue), normal lung cells and infiltrating immune cells (green), heart tissue and eosinophilic cells (orange), blood vessels and areas of hemorrhage (red). Scale bars, 2 mm. F Two selected ROIs on the H&E-stained lung section of the HDM-treated mouse shown in E. ROI#1 (left panel) demonstrates dense perivascular and peribronchial immune infiltrates with high numbers of macrophages (inset). ROI#2 (right panel) shows an AC with papillary morphology invading the bronchial lumen (inset) and stromal desmoplasia (indicated by asterisks). Scale bars, 2 mm (whole lungs), 0.2 mm (ROI#1 and 2), and 0.1 mm (ROI#1 and 2 insets). G Tumor multiplicity calculated on H&E-stained sections as shown in E upper panels. H Tumor area calculated on QuPath-pseudocolored images as shown in E lower panels. I Tumors on H&E-stained sections as shown in E upper panels were classified into three grades (Grade 1, AAH and EH; Grade 2, AD; Grade 3, AC) and each grade was expressed as a percentage of the total. Data are presented as mean ± SEM. Statistical significance was assessed by one-way (D, G, and H) or two-way (B and I) ANOVA with post hoc Bonferroni’s test, or log-rank Mantel-Cox test (C). n.s: non-significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001