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. 2021 Jun 26;69:103452. doi: 10.1016/j.ebiom.2021.103452

Figure 3.

Figure 3

YAP/TAZ expression is increased and correlated with more α-SMA+ cells in CD with fibrostenosis. (a) YAP/TAZ expression was detected in fibroblasts isolated from stenotic and nonstenotic areas of intestines from CD patients via real-time quantitative polymerase chain reaction. (b-c) Representative images of immunohistochemical staining showing the expression of YAP/TAZ in different layers of stenotic and nonstenotic intestinal tissue from CD patients (n=10). Scale bar = 20 μm. (d-e) Western blot analysis of YAP/TAZ expression in 3 paired samples of fibroblasts derived from CD patients with and without stenosis. (f) Intestinal tissues from CD patients with and without stenosis (n=10) were double-stained for YAP and α-SMA, and (g) YAP+ cells and (h) α-SMA+ cells were quantified. Quantifications were performed with the average values in 3 representative HPFs/sample. Scale bar = 20 μm (upper panels) and 10 μm (lower panels). (i) The number of YAP+ cells was evaluated by immunofluorescence staining and correlated with the number of α-SMA+ cells in CD patients with and without stenosis (n=10). (j) Intestinal tissues from CD patients with and without stenosis (n=10) were double-stained for TAZ and α-SMA, and (k) α-SMA+ cells and (l) TAZ + cells were quantified. Quantifications were performed with the average values in 5 representative HPFs/sample. Scale bar = 20 μm (upper panels) and 10 μm (lower panels). (m) The number of TAZ+ cells was evaluated and correlated with the number of α-SMA+ cells in CD patients with and without stenosis (n=10). (n-o) Representative images of immunofluorescence staining showing the nuclear localization of YAP/TAZ in fibroblasts isolated from stenotic regions. Scale bar = 20 μm. In all cases, the bars in the graphs represent mean ± S.D. Statistical analyses were performed using Student's t test for (a) paired and (d-h, k-l) unpaired samples, Pearson's correlation for (i, m) correlation analysis and a chi-square test for (n-o) cell quantification. Significant differences are shown by *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. CD, Crohn's disease; HPF, high-power field; Case 1, Case 2 and Case 3 represent three paired samples from three individual CD patients.