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. 2020 Nov 23;23(12):101851. doi: 10.1016/j.isci.2020.101851

Figure 7.

Figure 7

Inhibiting the TGF-β Modulates the Degree of ECM Density and Fiber Organization

(A) HCT-116-containing LX-2 organoid diameter was measured over 96 hr in culture. A TGF-β inhibitor (SB 431542, 10 μM) was added to the media containing TGF-β (10 ng/mL) at the indicated time points. Graphs represent mean ± s.e.m (n = 4).

(B) Organoid stiffness was measured over 96 hr in culture conditions using a rheometer to determine the young's modulus of the construct. Graphs represent mean ± s.e.m (n = 4).

(C–F) Collagen fiber architecture in organoids treated with TGF- β inhibitor 24hr after TGF-β addition was analyzed with segmentation software, CT-FIRE, to generate distributions of fiber angle (C), fiber angle variance (D), and box and whisker plots of fiber length (E), and width (F). MATLAB was used to generate graphs from extracted data. Box and whisker plots with Tukey formatting of pooled fibers performed in triplicate, representing 500–5000 fibers in total. (∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001)