Gel contraction assay

Cell contraction was evaluated using the Collagen Gel Culturing Kit (Nitta Gelatin, Inc., Osaka, Japan). Collagen type I, 10× MEM, and reconstitution buffer (pH 7.3) were mixed in a 7:1:1 ratio while cooling, and it was mixed with a cell suspension adjusted to 1 × 10⁷ cells/mL in a ratio of 9: 1. The mixture at a volume of 500 μL was dispensed into each well of 24-well plates and incubated at 37°C for 60 minutes. The gels were released from the wells and four gels were transferred to each 6 cm dish containing DMSO or GSK1016790A (10, 100 nM) and 5 mL Trabecular Meshwork Cell Medium. After 0, 12, 24, 48, and 72 hours, the dishes containing gels were photographed with a gel imaging device and the area of each gel was quantified with ImageJ software.