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. 2021 Mar 25;15(5):1507–1527. doi: 10.1002/1878-0261.12937

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© 2021 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Fig. 3 — Cancer‐associated fibroblast migration and expression of ITGA11, integrin α5, integrin β1, and COL11A1. Sub‐confluent fibroblasts from 16 pairs of control fibroblasts and CAFs were cultured, and (A) proteins were extracted and analyzed by western blot to detect (B) ITGA11, (C) COL11A1, (D) integrin α5, and (E) integrin β1 expression. (B‐E; n = 16, Wilcoxon) Vertical axis: expression of protein normalized to β‐actin. (F) After sub‐confluent fibroblasts, from 16 pairs of control fibroblasts and CAFs, were cultured, chemotactic activity toward fibronectin (20 µg·mL⁻¹) was measured. (F; n = 16, Mann–Whitney) Vertical axis: number of migrated cells per 5HPF. Filled symbols represent lung adenocarcinoma. Open symbols represent squamous cell lung cancer. (G) Comparison of CAFs and control fibroblasts from the same donor. (G; n = 16, Wilcoxon) Vertical axis: number of migrated cells per 5HPF. Filled symbols represent lung adenocarcinoma. Open symbols represent squamous cell lung cancer. (H) Relationship between fold CAF migration toward fibronectin (20 µg·mL⁻¹) divided by corresponding control fibroblast migration and fold CAF expression of ITGA11 divided by corresponding control fibroblast. Vertical axis: fold CAF migration compared to control fibroblasts. Horizontal axis: fold ITGA11 expression in CAFs compared to control fibroblasts. (H; n = 16, Spearman) (I) Relationship between fold CAF migration toward fibronectin (20 µg·mL⁻¹) divided by corresponding control fibroblast migration, and fold CAF expression of COL11A1 divided by corresponding control fibroblast. Vertical axis: fold CAF migration compared to control fibroblasts. Horizontal axis: fold COL11A1 expression in CAFs compared to control fibroblasts. (I; n = 16, Spearman) (J) After sub‐confluent patients’ control fibroblasts and CAFs were cultured, migration activity toward collagen type I (1 µg·mL⁻¹) was measured. (J; n = 9, Mann–Whitney) Vertical axis: number of migrated cells per 5HPF. Filled symbols represent lung adenocarcinoma. Open symbols represent squamous cell lung cancer. Each symbol represents one patient. The values represent the mean ± SD. Pts: patients. **P < 0.01, ***P < 0.001.