b-0702  (Echelon Biosciences)

Journal: PLoS ONE

Article Title: Non-Invasive Imaging of Tumors by Monitoring Autotaxin Activity Using an Enzyme-Activated Near-Infrared Fluorogenic Substrate

doi: 10.1371/journal.pone.0079065

Figure Lengend Snippet: ( a ) AR-2 was incubated with varying concentrations of purified ATX and fluorescence was measured over time. ( b ) AR-2 was incubated with enzymes homologous to ATX. Enzyme reactions included human and mouse ATX, human NPP1, human and mouse NPP7, shrimp alkaline phosphatase, human LPLA2 and human INPP4A. Mouse enzymes are indicated with (m). ( c ) AR-2 was incubated with media from COS-7 cells transfected with ATX-encoding or mock plasmid and fluorescence was monitored over time. ( d ) AR-2 was incubated with conditioned media from cancer cells overexpressing ATX in the presence and absence of PF-8380 (1 µM), and fluorescence was monitored over time. For all panels results are means ± SD from at least three experiments.

Article Snippet: Purified human ATX, INPP4A, LPLA2, PF-8380, S32826, BrP-LPA and FS-3 were obtained from Echelon Biosciences, Inc. (Salt Lake City, UT).

Techniques: Incubation, Purification, Fluorescence, Transfection, Plasmid Preparation

Journal: PLoS ONE

Article Title: Non-Invasive Imaging of Tumors by Monitoring Autotaxin Activity Using an Enzyme-Activated Near-Infrared Fluorogenic Substrate

doi: 10.1371/journal.pone.0079065

Figure Lengend Snippet: ( a ) Mice bearing MDA-MB-231 xenografts were treated with vehicle or the ATX inhibitor PF-8380. Twenty minutes later, AR-2 was injected i.v. , and the mice were subsequently imaged with NIRF. ( b ) Tumors from (a) were imaged ex vivo . ( c ) Ex vivo tumor fluorescence was normalized to total protein levels (*** denotes p<0.001).

Article Snippet: Purified human ATX, INPP4A, LPLA2, PF-8380, S32826, BrP-LPA and FS-3 were obtained from Echelon Biosciences, Inc. (Salt Lake City, UT).

Techniques: Injection, Ex Vivo, Fluorescence

Journal: Cell reports

Article Title: Heterogeneity of human anti-viral immunity shaped by virus, tissue, age, and sex

doi: 10.1016/j.celrep.2021.110071

Figure Lengend Snippet:

Article Snippet: HLA-B*0702[RPHERNGFTVL]-CMV pp65 , ProImmune , F308-4A-D (APC).

Techniques: Flow Cytometry, Recombinant, Staining, Sequencing, RNA Sequencing Assay, Expressing, Software

Journal: Cell reports

Article Title: Heterogeneity of human anti-viral immunity shaped by virus, tissue, age, and sex

doi: 10.1016/j.celrep.2021.110071

Figure Lengend Snippet:

Article Snippet: HLA-B*0702[TPRVTGGGAM]-CMV pp65 , ProImmune , F045-4A-D (APC).

Techniques: Flow Cytometry, Recombinant, Staining, Sequencing, RNA Sequencing Assay, Expressing, Software

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Disrupted Blood‐Brain Barrier and Mitochondrial Impairment by Autotaxin–Lysophosphatidic Acid Axis in Postischemic Stroke

doi: 10.1161/JAHA.121.021511

Figure Lengend Snippet: A , Enzymatic activity test for ATX, measured with AR‐2 fluorescence, quantified as relative fluorescence unit (RFU), in sham, ischemia and reperfusion (I/R), HA130, and PF8380 mouse brain slices. B , Lysophosphatidylcholine (LPC) subspecies in plasma from sham, I/R, HA130, and PF8380 mice measured with high‐performance liquid chromatography–tandem mass spectrometry (liquid chromatography–mass spectrometry [LC‐MS]) quantified relative to 18:1 LPC. C , LPA subspecies in plasma from sham, I/R, HA130, and PF8380 mice measured with LC‐MS, quantified relative to 18:1 LPA. D and E , Graph represents oxygen consumption rate (OCR) (pmol/min per µg protein) measurements in isolated mitochondria from sham, I/R, HA130, PF8380, or BrP‐LPA mouse brain tissue at baseline and after sequential addition of oligomycin, carbonyl cyanide p‐trifluoro‐methoxyphenyl hydrazone (FCCP), and antimycin A+rotenone. F , Spare respiratory capacity, ATP turnover, and maximum respiration values analyzed in sham, I/R, HA130, PF8380, or BrP‐LPA mice. All values are mean±SD. * P <0.05, ** P <0.01 (1‐way ANOVA, followed by the Tukey, post hoc test, was performed).

Article Snippet: The autotaxin inhibitors PF8380 (30 mg/kg body weight) (Echelon Biosciences Inc, Logan, UT), HA130 (0.5 mg/kg body weight; 1 μL/g, 1 mmol/L) (Echelon Biosciences Inc), and BrP‐LPA (10 mg/kg body weight) were injected intraperitoneally in mice 1 hour before MCAO surgery in the respective separate experimental groups., , The effect of LPA on vascular permeability was examined, as shown previously.

Techniques: Activity Assay, Fluorescence, High Performance Liquid Chromatography, Mass Spectrometry, Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Isolation

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Disrupted Blood‐Brain Barrier and Mitochondrial Impairment by Autotaxin–Lysophosphatidic Acid Axis in Postischemic Stroke

doi: 10.1161/JAHA.121.021511

Figure Lengend Snippet: A , Triphenyl tetrazolium chloride staining of mouse brain slices and respective infarct volume (percentage of the hemisphere) measured in sham, ischemia and reperfusion (I/R), HA130, PF8380, or BrP‐LPA mice (n=5 in each group). B , Superoxide measured using high‐performance liquid chromatography in brain tissue lysate after staining with a dihydroethidium fluorescence probe in sham, I/R, HA130, PF8380, or BrP‐LPA mice. C through E , Superoxide dismutase (SOD) activity, catalase activity, and glutathione peroxidase activity quantified in brain tissue lysate from sham, I/R, HA130, PF8380, or BrP‐LPA mice. F , Glutathione levels were measured in sham, I/R, HA130, PF8380, or BrP‐LPA mice. All values are mean±SD (1‐way ANOVA, followed by the Tukey, post hoc test, was performed).

Article Snippet: The autotaxin inhibitors PF8380 (30 mg/kg body weight) (Echelon Biosciences Inc, Logan, UT), HA130 (0.5 mg/kg body weight; 1 μL/g, 1 mmol/L) (Echelon Biosciences Inc), and BrP‐LPA (10 mg/kg body weight) were injected intraperitoneally in mice 1 hour before MCAO surgery in the respective separate experimental groups., , The effect of LPA on vascular permeability was examined, as shown previously.

Techniques: Staining, High Performance Liquid Chromatography, Fluorescence, Activity Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Disrupted Blood‐Brain Barrier and Mitochondrial Impairment by Autotaxin–Lysophosphatidic Acid Axis in Postischemic Stroke

doi: 10.1161/JAHA.121.021511

Figure Lengend Snippet: A , Evans blue fluorescence measured and quantified with relative fluorescence unit (RFU) in whole brains of sham, ischemia and reperfusion (I/R), HA130, PF8380, or BrP‐LPA mice. B , Immunofluorescence staining for claudin‐5 in ipsilateral cortex of sham, I/R, HA130, PF8380, or BrP‐LPA mice. C , Immunofluorescence staining for zonula occludens‐1 in ipsilateral cortex of sham, I/R, HA130, PF8380, or BrP‐LPA mice. Bar=200 µm. All values are mean±SD (1‐way ANOVA, followed by the Tukey, post hoc test, was performed). CD31 indicates cluster of differentiation 31; and DAPI, 4′,6‐diamidino‐2‐phenylindole.

Article Snippet: The autotaxin inhibitors PF8380 (30 mg/kg body weight) (Echelon Biosciences Inc, Logan, UT), HA130 (0.5 mg/kg body weight; 1 μL/g, 1 mmol/L) (Echelon Biosciences Inc), and BrP‐LPA (10 mg/kg body weight) were injected intraperitoneally in mice 1 hour before MCAO surgery in the respective separate experimental groups., , The effect of LPA on vascular permeability was examined, as shown previously.

Techniques: Fluorescence, Immunofluorescence, Staining