Journal:

Article Title: Lysophosphatidic acid (LPA) and Angiogenesis

doi: 10.1007/s10456-008-9113-5

Figure Lengend Snippet: To analyze the role of LPA receptors in LPA-induced angiogenesis we used the LPA1 and LPA3 receptor antagonist, VPC32183. Using the same approach aforementioned, we show in 3A and 3B that LPA-induced angiogenesis is blocked completely by this compound at 10µM. Importantly, VPC32183 alone (at 10µM) had no discernable effect in angiogenesis and does not block VEGF-induced angiogenesis (Figure 3C). One-Way ANOVA test was performed between groups. ** P< 0.001. Error bars represent standard deviation (SD). In figure 3B (scale bar: 0.125 cm), we present the image of a single membrane representative of each group.

Article Snippet: Disclosures : VPC32183 is sold by Avanti Polar Lipids under license from the University of Virginia.

Techniques: Blocking Assay, Standard Deviation, Membrane

Journal:

Article Title: Lysophosphatidic acid (LPA) and Angiogenesis

doi: 10.1007/s10456-008-9113-5

Figure Lengend Snippet: S-1-oleoyl-2-O-methylglycerophosphothionate (S-OMPT), at 100nM, is able to induce a strong angiogenic response, similar to the LPA response at this concentration (4A–4C). CAMs treated with S-OMPT (100nM) together with VPC32183 (10µM) document that the response is completely blocked by inclusion of this antagonist (4C). For statistical analysis, Student t test was performed between groups. ** P< 0.001. Error bars represent standard deviation (SD). Figure 4B is the image of a single membrane that is representative of each group. Scale bar: 0.125 cm.

Article Snippet: Disclosures : VPC32183 is sold by Avanti Polar Lipids under license from the University of Virginia.

Techniques: Concentration Assay, Standard Deviation, Membrane

Journal:

Article Title: Lysophosphatidic acid (LPA) and Angiogenesis

doi: 10.1007/s10456-008-9113-5

Figure Lengend Snippet: CAMs were treated with recombinant sphingomyelinase D (SMaseD) and the number of vessels that intersected the disc was counted. This enzyme is able to induce an angiogenic response (5A) similar to the LPA response. Two catalytically inactive forms of this enzyme (H37N and H73N) were unable to induce angiogenesis (5B), suggesting that the response obtained is dependent on the catalytic activity (lyso-PLD) of the arachnid enzyme. Moreover, SMaseD-induced angiogenesis is blocked by the LPA receptor antagonist VPC32183 at 10µM (5C), suggesting that the positive response obtained is dependent on the activation of LPA1,3 receptors. One-Way ANOVA test was performed between groups. ** P< 0.001. Error bars represent standard deviation (SD).

Article Snippet: Disclosures : VPC32183 is sold by Avanti Polar Lipids under license from the University of Virginia.

Techniques: Recombinant, Activity Assay, Activation Assay, Standard Deviation

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

Journal: Immunity

Article Title: Polyreactive Broadly Neutralizing B cells Are Selected to Provide Defense against Pandemic Threat Influenza Viruses

doi: 10.1016/j.immuni.2020.10.005

Figure Lengend Snippet:

Article Snippet: ProteOn Phosphoric Acid Solution (0.85%) , BioRad , Cat#1762260.

Techniques: Virus, Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Software