Figure 2

Impact of fibroblast growth factor receptor 3-IIIc (FGFR3-IIIc) on growth and clonogenicity. (A) SW480 and (B) HCT116 transfectants were plated at 3 × 10³ cells per well in 96-well plates and growth was determined by 3′ (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay at the indicated time points (A) or after 3 days (B). Data points represent the mean±s.e.m. of 10 determinations from two independent experiments (left panels). In all, 2.5 × 10⁴ cells were plated for assessment of DNA synthesis. After 24 h, cultures were incubated with 1 μCi ml^{–1 3}H-thymidine for 1 h and incorporation of radioactivity into DNA was determined as described in Sonvilla et al (2008) (right panels). (C) Stable SW480 and HCT116 transfectants were plated at 100 and 200 cells per well, respectively, in six-well plates, fixed and stained with crystal violet 10 days later to show colony formation. Colonies were counted from two experiments using duplicate cultures (means±s.e.m.). SW480 (left panel), HCT116 (right panel). (D) SW480 and Caco2 cells were infected with kinase-dead FGFR3 (KD3)-IIIcv or control virus (Cv). Cloning efficiency was determined from 200 cells per well and compared with untransduced control (UC). ^*, ^**, ^*** and #, ### indicate increase or a decrease, respectively, as compared with vector controls at P<0.05, P<0.01 or P<0.0001 (Student's t-test). d, day.