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. 2017 Jul 7;28(14):1871–1882. doi: 10.1091/mbc.E17-01-0014

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© 2017 Herum et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — Stretch promotes ECM remodeling by CFBs. (A) Schematic illustration of the principles of the stretch model. (B) Collagen (col) 1a1, 1a2, and 3a1 and fibronectin (FN1) mRNA normalized to GAPDH mRNA in CFBs on 3 and 8 kPa gels subjected to 0%, 3%, and 6% stretch (x-axis). One-way ANOVA was used to test significant effect of stretch with Tukey’s post hoc tests for specific comparisons. Statistical results are indicated in the figure. N = 4 (control) and 12 (stretch). (C) Quantification and immunoblot of FN protein normalized to total protein determined by Ponceau staining. N = 4 (3 kPa) and 8 for (8 kPa). (D) Fold difference in LOX, BMP-1, OPN, POSTN, TNC, SPARC, THBS1, matrix metalloproteinase (MMP) 2, and TIMP 1 and 2 mRNA expression normalized to GAPDH mRNA. Significance was determined by multiple Student’s t tests corrected for multiple comparisons using Holm-Sidak’s post hoc test. p Values displayed in D are the uncorrected p values from the t test. N = 4 (control) and 12 (stretch). (E) Relative proliferation of CFB controls (C) or with 3% stretch (S). N = 3. (F) SMA mRNA normalized to GAPDH mRNA. N = 4 (control) and 12 (stretch). (G) Immunofluorescence staining for SMA (green) and DAPI staining for nuclei (blue). (H) Immunoblot for total and phophorylated focal adhesion kinase (FAK and pFAK, respectively) normalized to GAPDH. N = 4. Student’s t test was used to determine significant changes for C, E, F, and H. *P < 0.05, **P < 0.01, ***P < 0.001; n.s., nonsignificant.