Sphingosine 1-phosphate (S1P) reduces hepatocyte growth factor-induced migration of hepatocellular carcinoma cells via S1P receptor 2
Fig 2
Effects of SB203580, SP600125, PD98059, deguelin or Y27632 on the HGF-induced migration, and phosphorylation of p38 MAPK (A), JNK (B), ERK (C), AKT (D) and MYPT-1 (E) in HuH7 cells. For the cell migration, the cells were pretreated with the indicated doses of SB203580 (A), SP600125 (B), PD98059 (C), deguelin (D) or Y27632 (E) for 60 min, and then stimulated by 30 ng/ml of HGF or vehicle for 23 h. The migrated cells were fixed with paraformaldehyde, and stained with DAPI for nucleus (blue signal). The cells were photographed by fluorescent microscopy at a magnification of 20× (upper panel) and counted (bar graph). For the Western blot analysis, the cells were pretreated with the indicated doses of SB203580 (A), SP600125 (B), PD98059 (C), deguelin (D) or Y27632 (E) for 60 min, and then stimulated by 30 ng/ml of HGF or vehicle for 5 min ((A), (B) and (E)) or 3 min ((C) and (D)). Each value represents the mean ± SD of triplicate determinations from three independent cell preparations. * p<0.05 compared to the value of the control cells without HGF. **p<0.05 compared to the value of the cells with HGF alone. N.S. designates no significant difference between the indicated pairs. Scale bar: 100 μm.