Fig. 1. Deposition of FN regulates the cellular migration area.

a Schematic representation of microenvironment remodelling. During collective cell migration, ECM is continuously degraded and constructed. b Immunostaining results of the nucleus, actin and FN. NHBE cells were initially placed in a triangular shape, and were observed to have migrated towards the lower right corner after 24 hours, although FN remained in the area without cells. c Schematics of the procedure for simultaneous dual patterning of cells and FN. d Phase contrast and immunostaining of the nucleus (DAPI), actin and FN show the dual patterning of NHBE cells and FN. A rectangular FN boundary was patterned on a dish, and NHBE cells then patterned in the centre of the FN area. e Tracking of cell migration inside the FN-patterned area. The red dotted line indicates the FN boundary, which was determined from the staining results. f Cellular behaviour in the crowded area near the FN boundary. Immunostaining of FN is shown in red. g Control experiment without FN patterning. The complete time-lapse movies for parts e–g are shown in Supplementary Movies 1–3. Scale bar: 100 μm (b, e–g) and 1 mm (d).