In immunofluorescence staining, cells seeded in a 35-mm glass-bottom dish (Matsunami Glass Ind., Ltd., Osaka, Japan) were stained with anti-mouse fibronectin antibody (1:50, BD Biosciences US, CA, USA) or anti-mouse collagen antibody (1:50, Novus Biologicals USA, CO, USA) diluted in a 1:1 mixture of PBS and media (200 μl) for 30 min at 4°C, washed twice with the mixture solution and stained with anti-mouse fluorescein isothiocyanate–conjugated IgG (1:100, Jackson ImmunoResearch Laboratory Inc., Pennsylvania, USA) or anti-rabbit rhodamine-conjugated IgG (1:100, Jackson ImmunoResearch Laboratory Inc.), respectively, diluted in a 1:1 mixture of PBS and media (200 μl) for 30 min at 4°C. After washing twice, the holder was observed under a fluorescence optical microscope.

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