Cell migration was determined using the wound healing scratch assay as previously described [5] (link), [26] (link). Cells were seeded on a 3.5 cm dish and grown overnight. Confluent cells were cultured in DMEM containing 0.5% FBS treated with 5 µg/mL mitomycin-C. After 3 days incubation in LG or HG media, cells were transferred to the medium containing low FBS (0.5%) and 5 µg/mL mitomycin-C. Effects of HG, bFGF (100 ng/mL) and FGFR1 inhibitor PD173074 (50 nM) on wound healing were measured after 12, 24, 36 and 48 hours. Images of the wounded cell monolayers were taken using a microscope (model IX70; Olympus, Tokyo, Japan) at 0, 12, 24, 36 and 48 hours after wounding and recorded for 48 hours using a microscope (model IX-70; Olympus) equipped with a CCD Camera (CoolSNAP HQ; Nippon Roper, Chiba, Japan) and controlled by MetaMorph software (Universal Imaging Co., Ltd., UK). All experiments were performed in the presence of 5 µg/mL of mitomycin-C to inhibit cell proliferation.
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