Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Vehicle-dependent Effects of Sphingosine 1-phosphate on Plasminogen Activator Inhibitor-1 Expression

doi: 10.5551/jat.37663

Figure Lengend Snippet: S1P bound to albumin induced PAI-1 expression by activating S1PR2 pathway in 3T3L1 adipocytes. (A) 3T3L1 adipocytes were pre-incubated for 18 hours in FBS-free DMEM and the medium was exchanged for FBS-free DMEM containing 20 µM of VPC23019, JTE013, or DMSO for 30 minutes. Then, the cells were challenged with a medium containing 10 µM of sphingosine 1-phosphate (S1P) with the corresponding inhibitors for 4 hours. The mRNA level of plasminogen activator inhibitor 1 (PAI-1) was analyzed using real-time PCR. 18S was utilized as an internal control ( n = 4–6/group). † p < 0.01 vs. DMSO alone, YC1 alone, S1P + VPC, and JTE alone, and p < 0.05 vs. S1P + JTE. ‡ p < 0.01 vs. other groups. (B, C). 3T3L1 adipocytes were pre-incubated for 18 hours in FBS-free DMEM, and the medium was exchanged for FBS-free DMEM with 10 µM of S1P bound to apoM-rich vehicle (ApoM) or control vehicle (Null) for 4 hours. Then, the modulation of the phosphorylation of moesin in 3T3L1 cells was examined using western blotting ( n = 6/group). ** p < 0.01, *** p < 0.001.

Article Snippet: VPC23019 (857360P; Avanti Polar Lipids, Alabaster, AL), JTE013 (10009458; Cayman Chemical, Ann Arbor, MI), Y27632 (257-00511; WAKO Pure Chemical Industries, Osaka, Japan), wortmannin, YC-1 (W1628, Y102; Sigma-Aldrich Co), and SIS3 and BAY11-7082 (sc-222318, sc-200615; Santa Cruz Biotechnology, Inc. TX) were dissolved in DMSO.

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Control, Phospho-proteomics, Western Blot

Journal: Cancer Research

Article Title: Growth of Triple-Negative Breast Cancer Cells Relies upon Coordinate Autocrine Expression of the Proinflammatory Cytokines IL-6 and IL-8

doi: 10.1158/0008-5472.can-12-4524-t

Figure Lengend Snippet: Figure 3. LPA induction of NF-kB activation is critical for IL-6, IL-8, and CXCL1 expression and is dependent upon high EZH2 expression in TNBC cells. qRT- PCR determination of: IL-6 (A), IL-8 (B), and CXCL1 (C) RNA expression in SUM159 cells after 24 hours of LPA-stimulation (5 mmol/L; n ¼ 3). D, induction of transcription factors determined by dual luciferase assays in LPA-stimulated (5 mmol/L) SUM159 cells. E and F, ELISA determination of IL-6, IL-8, and CXCL1 protein expression in SUM159 cells after 24 hours of NF-kB inhibition (10 mmol/L; n ¼ 6; E) or treated with LPA (5 mmol/L) and inhibitors (F). G, qRT-PCR expression of EZH2 in triple-negative/basal-like and ER-positive/luminal-like breast cancer cell lines (n ¼ 4). H, ELISA determination of IL-6, IL-8, and CXCL1 in SUM159 cells after treatment with siEZH2 (n ¼ 3). I, colony formation with EZH2 inhibition in SUM159 cells (n ¼ 4). Statistically significant inhibition compared with control is denoted with , P < 0.05; , P < 0.01.

Article Snippet: Lysophosphatidic Acid (LPA 18:1) and LPA inhibitors (VPC32183 and VPC12249) were obtained from Avanti Polar Lipids. siRNA transfection EZH2 siRNAs andLPAR2were purchased fromSigma. siRNA transfection was conducted using a final concentration of Authors' Affiliations: Departments of 1Clinical Cancer Prevention and 2Systems Biology, The University of Texas MD Anderson Cancer Center; and 3Lester and Sue Smith Breast Center, Department of Medicine, Baylor College of Medicine, Houston, Texas Note: Supplementary data for this article are available at Cancer Research Online (http://cancerres.aacrjournals.org/).

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, RNA Expression, Luciferase, Enzyme-linked Immunosorbent Assay, Inhibition, Control