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. 2022 Aug 30;113(11):3710–3721. doi: 10.1111/cas.15483

FIGURE 5.

FIGURE 5

Effect of COX‐2 inhibitor treatment on dome‐like structures. (A) Immunofluorescence images of COX‐2 in bronchial epithelia from CK19‐YFP or CK19‐RasV12 mice with tamoxifen treatment. Scale bars, 20 μm. (B) Quantification of COX‐2‐positive YFP or RasV12 cells. Data are mean ± SEM n = 183 (not extruded in CK19‐YFP), 79 (not extruded in CK19‐RasV12), and 231 (dome‐like structure in CK19‐RasV12) cells from three (CK19‐YFP and not extruded in CK19‐RasV12) or four (dome‐like structure in CK19‐RasV12) mice. ***p < 0.005, one‐way anova with Tukey's test. (C) Experimental design for the ibuprofen treatment in CK19‐RasV12 mice. CK19‐RasV12 mice were continuously administrated with ibuprofen in drinking water starting at 3 days before tamoxifen injection and analyzed at 2 weeks. (D, E) Quantification of the effect of ibuprofen on the number (D) and length (E) of dome‐like structures. (D) Data are mean ± SEM n = 12 sections from four mice. (E) Data are median ± quartiles. n = 109 (water) and 89 (ibuprofen) dome‐like structures from three mice. *p < 0.05, unpaired two‐tailed Student's t‐test (D) or NS, not significant, Mann–Whitney test (E). (F) Immunofluorescence images of α‐SMA in bronchial epithelia from CK19‐RasV12 mice with ibuprofen and tamoxifen treatment. Arrowheads indicate α‐SMA‐positive activated fibroblasts within the dome‐like structure, whereas arrows indicate α‐SMA‐positive smooth muscle underneath the epithelium. Note that smooth muscle has much higher α‐SMA expression compared with activated fibroblasts. Scale bars, 20 μm. (G) Quantification of the effect of ibuprofen on α‐SMA‐positive activated fibroblasts in dome‐like structures. Data are mean ± SEM n = 105 (water) and 129 (ibuprofen) dome‐like structures from five mice. ***p < 0.005, unpaired two‐tailed Student's t‐test. (H, I) Quantification of the effect of ibuprofen on Ki‐67+ surrounding epithelial cells (H) or CD45+ immune cells (I) in dome‐like structures. (H) Data are mean ± SEM n = 1575 (water) and 1301 (ibuprofen) cells from three mice. (I) n = 70 (water) and 64 (ibuprofen) dome‐like structures from three mice. NS, not significant, unpaired two‐tailed Student's t‐test.