P mek 2338s

S1P was obtained from Avanti Polar Lipid Inc. (Alabaster, AL, USA). We obtained SphK1 (GTX33516) antibody from Genetex (Irvine, CA, USA), Ras (SC-520), Raf (SC-133), MEK (SC-6250), ERK (SC-1647), PDGF-A (SC-9974), c-Jun (SC-74543), p-Raf (SC-101791), p-ERK (SC-7383) and p-c-Jun (SC-822) from Santa Cruz (Santa Cruz, CA, USA), and p-MEK (2338S) from Cell Signaling Technology (Danvers, MA, USA). ON-TARGETplus siRNAs were purchased from Dharmacon Research (Lafayette, CO, USA). SphK1 cDNA clone plasmid was purchased from OriGene (Rockville, MD, USA). Gibco-BRL Life Technologies (Grand Island, NY, USA) supplied fetal bovine serum (FBS), DMEM, α-MEM, and all other cell culture reagents. Promega (Madison, WI, USA) supplied the pSV-β-galactosidase vector and luciferase assay kits. The AP-1 luciferase plasmid was purchased from Stratagene (La Jolla, CA). All other chemicals or inhibitors were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Anti-mouse monoclonal antibodies specific for MEK (sc-6252), p-ERK (sc-7383), ERK (sc-1647), p-STAT3 (sc-56747), CCL4 (sc-130330), and anti-rabbit monoclonal antibodies specific for STAT3 (sc-482) were purchased from Santa Cruz Biotechnology (Shanghai) Co., Ltd. (Pudong New District, Shanghai, China). CD31 (ab9498) and Angpt2 (ab153934) antibodies were purchased from Abcam (Cambridge, UK). P-MEK (2338s) was purchased from Cell Signaling Technology, Inc. (Taigen Bioscience Corporation, Taipei, Taiwan). Beta-actin antibody (GT5512) was purchased from GeneTex, Inc. (Irvine, CA, USA). The control, MEK, ERK and STAT3 small interfering RNAs (siRNAs) were purchased from GE Healthcare Dharmacon, Inc. (Lafayette, CO, USA). Invitrogen™ Lipofectamine™ 2000 was purchased from Thermo Fisher Scientific (Carlsbad, CA, USA). Human CCL4 recombinant protein was purchased from PeproTech US (Cranbury, NJ, USA). Matrigel was purchased from Corning Life Sciences (Tewksbury, MA, USA). All other chemicals were purchased from Merck SA (Darmstadt, Germany).