Fig. 3. The MEK/ERK pathway is involved in AR-induced VEGF-C expression and lymphangiogenesis in human chondrosarcoma cells. Cells were pretreated with U0126 and PD98059 for 30 min or transfected with siRNAs for 24 h, then stimulated with AR (50 μg/ml) for 24 h. VEGF-C expression was examined by qPCR (A) and ELISA (B). The medium was collected as CM and then applied to the LECs. Capillary-like structure formation and LEC migration was examined by tube formation and the Transwell assay, respectively (C and D). (E) JJ012 cells were incubated with AR (50 μg/ml) for 120 min, and MEK/ERK phosphorylation was determined by Western blotting. (F) JJ012 cells were pretreated with inhibitors as indicated, then incubated with AR for 60 min and analyzed by Western blotting with MEK and ERK antibodies. Western blot data were normalized to MEK and ERK as the loading control. Results are expressed as the mean SEM. *P<0.05 compared with control; #P<0.05 compared with the AR-treated group.