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Fig. 6. Dual imaging of fibronectin and LTBP1 in a day-12 FRC culture. (A) Still image frames taken at the start of time-lapse imaging in a day 12 FRC culture using fluorescent probes for fibronectin (green) and LTBP1 (red). Note that the LTBP1 and fibronectin fibrils are localized in separate fibrillar networks. Movie 6 in supplementary material shows that both fibrillar networks undergo continual stretching and contraction in response to cell movement. (B) Time-lapse image series from the boxed area in A. Note a fibronectin fibril (large arrow) that stretches between 15 minutes and 4 hours, then contracts between 4 and 8 hours. The fibril then stretches again to reach its maximal length at 12 hours, then contracts again between 12 and 18 hours. Also note an LTBP1 fibril (small arrow) in the same region that appears to show much less motion compared with the fibronectin fibril. The bottom panel shows the trajectories of these fibrils obtained by tracking the co-ordinates of the end of each fibril in the image series and confirms that the LTBP1 fibril traverses a smaller trajectory. See also Movie 7 in supplementary material. Bar, 100 µm (A); 50 µm (B).

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