-fold and 2.53 0.13-fold higher, respectively, with LPA treatment as compared to those with vehicle treatment. LPA treatment enhances cytokine C5/C5a, PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19764249 M-CSF, MCP-5, and SDF-1 Debio-1347 cost expression in liver sinusoidal endothelial cells Conditioned media derived from vehicle treated and LPA treated liver sinusoidal endothelial cells were used to determine cytokines’ expression using a cytokine protein array. This showed that the spots for C5/C5a, M-CSF, MCP-5, and SDF-1 had different intensities between 5 / 13 LPA Effects on Liver Sinusoidal Endothelial Cells Fig 2. LPA effects on liver sinusoidal endothelial cells angiogenesis related protein expression. Conditioned media derived from vehicle and LPA treated liver sinusoidal endothelial cells were applied to angiogenesis protein array analysis. Data shown are representative images of three independent experiments. Significantly changed protein spots are indicated. Quantitative results for angiogenesis protein arrays. Results are the ratios of LPA treatment versus vehicle treatment. , ratio of > 2 was defined as a significant change. doi:10.1371/journal.pone.0122060.g002 conditioned media after vehicle treatment and after LPA treatment. Quantitative results showed that LPA treatment had enhanced C5/C5a, M-CSF, MCP-5, and SDF-1 PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19763407 expression relative to those after vehicle treatment. Fig 3. LPA effects on liver sinusoidal endothelial cells cytokine expression. Conditioned media derived from vehicle and LPA treated liver sinusoidal endothelial cells were applied to cytokine protein array analysis. Data shown are representative images of three independent experiments. Significantly changed protein spots are indicated. Quantitative results for cytokine protein arrays. Results are the ratios of LPA treatment versus vehicle treatment. , ratio of > 2 was defined as a significant change. doi:10.1371/journal.pone.0122060.g003 6 / 13 LPA Effects on Liver Sinusoidal Endothelial Cells Fig 4. LPA effects on liver sinusoidal endothelial cells chemokine expression. Conditioned media derived from vehicle and LPA treated liver sinusoidal endothelial cells were applied to chemokine protein array analysis. Data shown are representative images of three independent experiments. Significantly changed protein spots are indicated. Quantitative results for chemokine protein arrays. Results are the ratios of LPA treatment versus vehicle treatment. , ratio of > 2 was defined as a significant change. doi:10.1371/journal.pone.0122060.g004 LPA treatment enhances chemokine MCP-5, gp130, CCL28, and CXCL16 expression in liver sinusoidal endothelial cells Different conditioned media were also used to determine chemokines’ expression using a chemokine protein array. This showed that the spots for MCP-5, gp130, CCL28, and CXCL16 had different intensities between conditioned media after vehicle treatment and after LPA treatment. Quantitative 
results showed that LPA enhanced MCP-5, gp130, CCL28, and CXCL16 expression relative to those after vehicle treatment. LPA induced angiogenesis factor, cytokine, and chemokine expression in liver sinusoidal endothelial cells is mediated primarily through LPAR1 and LPAR3 signaling LPA regulates several proteins based on the LPAR subtype. Our results showed that LPAR1 and LPAR3 mRNA’s were strongly expressed in liver sinusoidal endothelial cells after LPA treatment. Thus, we investigated LPAR1 and LPAR3 involvement in LPA-mediated angiogenesis factor, cytokine, and chemokine expression in liver sinusoid