Interleukin-1β Expression Is Required for Lysophosphatidic Acid-Induced Lymphangiogenesis in Human Umbilical Vein Endothelial Cells

Figure 1

Lysophosphatidic acid- (LPA-) induced lymphatic marker expressions are mediated through endothelial growth factor receptor (EGFR) transactivation-, matrix metalloproteinase- (MMP-), LPA1/3-, and interleukin- (IL-) 1R-dependent pathways in human umbilical vein endothelial cells (HUVECs). Starved HUVECs were pretreated with AG1478 (100 nM), AF12198 (10 nM), GM6001 (10 𝜇M), Ki16425 (10 𝜇M), and toxin B (10 nM) for 1 h, followed by LPA (5 𝜇M) treatment for an additional 8 h. HUVECs were dissociated by trypsinization and fixed with 4% paraformaldehyde. Fixed cells were labeled with antibodies for Prox-1, LYVE-1, and podoplanin, and then analyzed by CyFlow. Cells were also stained with a normal mouse or goat immunoglobulin G antibody, which served as a negative control.