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. 2013 Feb 4;34(3):360–366. doi: 10.1038/aps.2012.186

Figure 2.

Figure 2

Sphingosine-1-phosphate (S1P) upregulates vascular endothelial growth factor (VEGF)-C expression by a matrix metalloproteinase (MMP)-2-dependent pathway in human umbilical vein endothelial cells (HUVECs). (A) HUVECs were pretreated with GM6001 (10 μmol/L) for 1 h, followed by S1P (5 μmol/L) treatment for another 4 h. VEGF-C mRNA expression levels were monitored by real-time PCR. (B) siRNA mediated downregulation of MMP-2 mRNA expression in HUVECs. MMP-2 and scrambled siRNA were transfected into HUVECs by electroporation. The transfection efficiency was evaluated by real-time PCR. MMP-2 expression was suppressed in HUVECs transiently transfected with MMP-2 siRNA. (C) HUVECs transfected with scrambled or MMP-2 siRNA were incubated with S1P (5 μmol/L) for 4 h, and VEGF-C mRNA expression levels were monitored by real-time PCR. (D) HUVECs were incubated with S1P (5 μmol/L) for 8 h, and the secreted concentration of VEGF-C protein was determined by ELISA. Quantified results are shown as the mean±SD from at least three independent experiments. bP<0.05, cP<0.01 vs control.