Figure 3

Induction of apoptosis by dominant-negative fibroblast growth factor receptor 3 (FGFR3). SW480 (upper panels) and Caco2 (lower panels) cells were infected with adenoviral constructs expressing kinase-dead FGFR3-IIIc (KD3-IIIcv) and control virus (Cv) at multiplicity of infections (MOIs) of 1 (SW480) or 20 (Caco2). (A) Cell viability was determined 6 days later by 3′ (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay. The data points represent the mean±s.e.m. of two independent experiments using quadruplet cultures. (B) Apoptotic cells were detected after staining with Hoechst dye and propidium iodide to visualise nuclear morphology and counted from 1000 cells, each in triplicate cultures. (C) To determine caspase activation, cell lysates were analysed by western blot using a monoclonal antibody recognising procaspase 3. Inserts depict representative blots. Quantification of band intensities was performed from three independent experiments. (D) Caspase activity was determined using a substrate predominantly cleaved by caspase 3. The data points represent the mean±s.e.m. of two independent experiments using quadruplet cultures. ^*, ^**Indicate increase as compared with Cv at P<0.05 and P<0.01, respectively. #, ## indicate decrease as compared with Cv at P<0.05 and P<0.01 (Student's t-test).