Figure 1

Knockdown of RON inhibits cell growth and clonogenicity. (a) Expression level of RON-β and MET protein from normal pancreatic ductal cell line (HPNE) and PDAC cell lines were analyzed by western blotting. (b) RON-β and MET expression levels were analyzed by western blotting (left) and quantitative RT–PCR (right) in RON knockdown clones (shRON cl.1 and shRON cl.2) and vector control cells. (c) 1000 cells per well were seeded in 96-well plates and cell proliferation rates were determined by MTT assay at indicated time points. Bars represent s.d. of six replicates. (d) Clonogenicity assays were performed as described previously.⁵⁰ Briefly, cells were seeded at 50, 250 and 500 cells per 6 cm² dish and cultured for 2 weeks. The cells were stained with crystal violet and colonies were counted. The data were presented as mean±s.d. of experiments performed in triplicates. *P<0.05 and **P<0.01 compared with their vector controls, respectively. (e) 3 × 10⁴/well cells were seeded in 24-well Matrigel invasion chambers and treated with 200 ng/ml of MSP for 24 h. Invaded cells were counted under microscope and the data represent mean±s.d. from triplicate experiments. *P<0.05 and **P<0.01 compared with their vector controls, respectively.