Journal: bioRxiv

Article Title: Fatty acid binding proteins shape the cellular response to activation of the glucocorticoid receptor

doi: 10.1101/2021.07.02.450968

Figure Lengend Snippet: GRE-transcriptional activity of GR in the absence or presence of FABP1, FABP2, FABP3, FABP4 or FABP5 in response to vehicle control or increasing concentrations of (A) hydrocortisone (B) dexamethasone or (C) prednisolone, was assessed using a reporter gene in COS-7 cells after 24 h treatment (n=3). (D) Expression of the GR in naïve COS-7 cells or following transfection with GR determined by immunoblotting. Transcriptional activity of the endogenous or transfected GR in the presence of FABP4 in response to (E) dexamethasone or (F) prednisolone (n=3). (G) FABP translocation to the nucleus in COS-7 cells transfected with GR and GFP-FABP1 or GFP-FABP4 following 24 h treatment with vehicle or an EC 80 concentration of hydrocortisone, dexamethasone or prednisolone (n=3). Data are mean ± SEM from n independent experiments, as stated. For concentration-response curves, symbols show means and error bars, S.E.M. *** p<0.001, two-way ANOVA with Sidak’s multiple comparison test. For bar graphs, bars show the mean, error bars the S.E.M. and symbols show the independent data points for each experiment. *** p<0.001, two-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: Immunoblotting used primary antibodies recognising FABP1 (Abcam (AB76812; rabbit, 1:300), FABP2 (gift from Dr Satoshi Kaiura, Dainippon Sumi-tomo Pharma Co. Ltd., Osaka, Japan; mouse; 1:400), FABP3 (R&D Systems MAB1678; rabbit, 1:200), FABP4 (Abcam AB92501; rabbit, 1:1000), FABP5 (Abcam AB37267; rabbit, 1:200), GR (Abcam AB183127; rabbit, 1:400) or β-actin (Cell Signaling Technology 3700S; mouse, 1:5000).

Techniques: Activity Assay, Control, Expressing, Transfection, Western Blot, Translocation Assay, Concentration Assay, Comparison

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Protective Effects of Dexmedetomidine and Oxycodone in Patients Undergoing Limb Ischemia-Reperfusion

doi: 10.12659/MSM.918261

Figure Lengend Snippet: Perioperative variables of I/R and immune parameters.

Article Snippet: For quantification the serum concentration by enzyme-linked immunosorbent assay (ELISA), commercially available ELISA kits for measuring human tumor necrosis factor-α (TNF-α, Proteintech), interleukin-6 (IL-6, Proteintech), brain-derived neurotrophic factor (BDNF, Proteintech), fatty acid binding protein 3 (FABP3, Elabscience), and endothelin-1 (ET-1, Elabscience) were used.

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Protective Effects of Dexmedetomidine and Oxycodone in Patients Undergoing Limb Ischemia-Reperfusion

doi: 10.12659/MSM.918261

Figure Lengend Snippet: Plasma FABP3 ( A ), ET-1 ( B ), and BDNF ( C ) levels. The serum concentration of fatty acid binding protein 3 (FABP3), endothelin-1 (ET-1), and brain-derived neurotrophic factor (BDNF) were measured before anesthesia (T0) and at 30 min (T8) and 6 h after tourniquet release (T9). Sham – control (ischemia-reperfusion, I/R) group; Dex – dexmedetomidine group; Oxy – oxycodone group. Data are expressed as mean±SD. + P<0.05 vs. T0; * P<0.05 vs. sham; # P<0.05 vs. Dex.

Article Snippet: For quantification the serum concentration by enzyme-linked immunosorbent assay (ELISA), commercially available ELISA kits for measuring human tumor necrosis factor-α (TNF-α, Proteintech), interleukin-6 (IL-6, Proteintech), brain-derived neurotrophic factor (BDNF, Proteintech), fatty acid binding protein 3 (FABP3, Elabscience), and endothelin-1 (ET-1, Elabscience) were used.

Techniques: Concentration Assay, Binding Assay, Derivative Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: ( A ) Structure of heart-type fatty acid-binding protein (FABP3, PDB ID 3WVM ) in complex with palmitate (C16:0). Protein structure is represented as ribbons and colored according to the secondary structure elements as follows: α-helixes shown in orange, β-sheets in cyan, and loops in gray. C16:0 is shown as dark blue sticks, while residues involved in ligand and water cluster binding are shown as light gray balls and sticks. Water molecules (red spheres) and hydrogen bonds (yellow dashed lines) are shown. ( B ) Human FABP3 amino acid sequence. Residues colored in yellow were not assigned from the NMR spectra either for the apo-form or for the holo-form, while residues in cyan were not assigned only for complexes of FABP3 with acylcarnitines (ACs). Crosspeaks for residues colored in gray were not observed in any 2D 1 H- N HSQC spectra. The black arrow points to the position of the TEV cleavage site. ( C ) Assigned 2D 1 H- N HSQC spectra of human apo-FABP3 in 20 mM K 2 HPO 4 /KH 2 PO 4 , 50 mM KCl buffer pH 7.6 (KPi). His-tagged FABP3 (noncleaved) is shown in blue, and cleaved FABP3 is shown in red. Backbone amide resonances are denoted as one letter symbol and residue number according to the sequence in B. Labels for the residues from the His-tag are shown in light blue. Side chain amide resonances were not assigned. Residues are numbered according to UniProt ID P05413.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay, Sequencing, Residue

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Toxicity of C16:0-carnitine in PANC-1 cells after 4 h of incubation in the presence or in the absence of 60 μM heart-type fatty acid-binding protein (FABP3) in the cell media. Data are shown as the mean ± SEM of 3 independent experiments in at least 6 technical replicates. * indicates a significant difference compared to the cells not subjected to FABP3 treatment at the respective C16:0-carnitine concentration.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Incubation, Binding Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Toxicity of C16:0-carnitine in native and heart-type fatty acid-binding protein (FABP3)-overexpressing PANC-1 cells after 4 h of incubation. Data are shown as the mean ± SEM of 3 independent experiments in at least 6 technical replicates. * indicates a significant difference compared to the native cells at the respective concentration of C16:0-carnitine.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay, Incubation, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Superposition of the 2D 1 H- 15 N HSQC spectra for ( A ) heart-type fatty acid-binding protein (FABP3)-C18:1(n-9) t and ( B ) FABP3-C18:1(n-9) t -carnitine complex in 20 mM K 2 HPO 4 /KH 2 PO 4 , 50 mM KCl buffer pH 7.6 (KPi). The spectrum of apo-FABP3—in blue, spectrum of the FABP3-ligand complex—in red. Residues with chemical shift perturbations (CSPs) larger than the mean plus one standard deviation are assigned and black arrows show the shift of the corresponding crosspeak. * marks the crosspeaks that have disappeared upon binding of the acylcarnitines (ACs).

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Mapping of the chemical shift perturbations (CSPs) caused by binding of ( A ) C8:0, ( B ) C12:0, ( C ) C14:0, ( D ) C16:0, ( E ) C18:1(n-9) c , ( F ) C20:5(n-3) c , ( G ) C8:0-carnitine, ( H ) C12:0-carnitine, ( I ) C14:0-carnitine, ( J ) C16:0-carnitine, ( K ) C18:1(n-9) c -carnitine, or ( L ) C20:5(n-3) c -carnitine onto the heart-type fatty acid-binding protein (FABP3) structure. The CSPs are color-coded, in which red indicates larger shifts, while blue indicates no changes in the averaged δ H and δ N NMR chemical shifts. Unassigned or disappeared residues are colored in gray. FABP3 structure is taken from PDB ID 3WVM.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: ITC results of the heart-type fatty acid-binding protein (FABP3) interaction with fatty acids (FAs) and acylcarnitines (ACs) in 20 mM K 2 HPO 4 /KH 2 PO 4 , 50 mM KCl buffer pH 7.6 (KPi) at 25 °C.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: ( A ) Superposition of the ITC titration curves of the heart-type fatty acid-binding protein (FABP3) interaction with C18:1(n-9) c in black and C18:1(n-9) c -carnitine in cyan. Both experiments were performed in 20 mM K 2 HPO 4 /KH 2 PO 4 , 50 mM KCl buffer pH 7.6 (KPi) at 25 °C. Graphical representation of the thermodynamic binding parameters of the FABP3 interaction with ( B ) C18:1(n-9) c and ( C ) C18:1(n-9) c -carnitine.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Titration, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Superposition of the ITC thermograms for heart-type fatty acid-binding protein (FABP3) titrated with ( A ) C18:1(n-9) c and ( B ) C18:1(n-9) c -carnitine at three different temperatures: 16 (cyan), 25 (black), and 37 °C (red).

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Heat capacity change, Δ C p , and temperature dependence of the binding free energy, Δ G . ( A ) Δ C p values for four fatty acids (FAs) and corresponding acylcarnitines (ACs) binding to heart-type fatty acid-binding protein (FABP3). ( B ) Δ G of FA or AC binding to FABP3 within the temperature range from 0 to 100 °C.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Comparison of the changes in the heat capacity, Δ C p , for the heart-type fatty acid-binding protein (FABP3)—ligand complexes.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Comparison

Journal: International Journal of Molecular Sciences

Article Title: Heart-Type Fatty Acid Binding Protein Binds Long-Chain Acylcarnitines and Protects against Lipotoxicity

doi: 10.3390/ijms24065528

Figure Lengend Snippet: Top. Competitive binding studies of heart-type fatty acid-binding protein (FABP3)-C18:1(n-9) t -carnitine and three fatty acids (FAs) of different chain lengths. ( A ) Reference titration of C18:1(n-9) t -carnitine to apo-FABP3. ( B ) FABP3-C8:0 complex titrated with C18:1(n-9) t -carnitine. ( C ) FABP3-C10:0 complex titrated with C18:1(n-9) t -carnitine. ( D ) FABP3-C18:1(n-9) t -carnitine complex titrated with C10:0. ( E ) FABP3-C18:1(n-9) t -carnitine complex titrated with C12:0. All experiments were performed in 20 mM K 2 HPO 4 /KH 2 PO 4 and 50 mM KCl buffer pH 7.6 (KPi) at 25 °C. Bottom. Schematic representation of ligand binding and competition. Protein is represented as the dark blue sector, acylcarnitines (ACs) as orange, and FAs as cyan circles. Green arrows indicate that the competition event between ligands was successful.

Article Snippet: The commercially available FuGENE ® HD transfection reagent (Promega Corporation, Madison, WI, USA) was mixed with the FABP3 (NM_010174) mouse tagged ORF clone plasmid (OriGene Technologies, Inc., Rockville, MD, USA) in a 4:1 ratio for transfection according to the manufacturer’s protocol.

Techniques: Binding Assay, Titration, Ligand Binding Assay

Journal: Journal of Medical Biochemistry

Article Title: Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background Fluorescence Quenching Immune Chromatography

doi: 10.2478/jomb-2018-0017

Figure Lengend Snippet: Schematic diagram of plasma panel test for H-FABP a. Small cup with bottom coated with gold colloid-conjugated antibodies; b. sample pad to which the sample plasma is added; c. test line (T-line), coated with capture antibody; d. control line (C-line); e. nitrocellulose membrane, on which background fluorescence for T-line and C-line is read; and f. absorbent pad that absorbs excess sample solution.

Article Snippet: Anti-human H-FABP antibody (FABP3 2302 SPTN-5, 5.1 mg/mL) and anti-human H-FABP antibody (FABP3 2304 SPTN-5, 5.0 mg/mL) were obtained from Medix Biochemica.

Techniques: Clinical Proteomics, Control, Membrane, Fluorescence

Journal: Journal of Medical Biochemistry

Article Title: Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background Fluorescence Quenching Immune Chromatography

doi: 10.2478/jomb-2018-0017

Figure Lengend Snippet: Background fluorescence quenching with varying amounts of immobilized capture antibody on the T-line and a series of H-FABP/gold colloid-conjugated antibody complex concentrations. H-FABP antibodies were immobilized on test cards from solutions of varying concentrations (0.5, 0.75, 1.0, or 1.5 mg/mL) for testing to determine the optimal capture concentration. Briefly, serial H-FABP/gold colloid-conjugated antibody complex solutions were prepared by mixing 3, 5, or 7 mL of 1.0 mg/mL H-FABP solution with gold colloid-conjugated antibody solution prepared as described in the »Materials and Methods«. The H-FABP capture solution was diluted in PBS to 0.5, 0.75, 1.0, or 1.5 mg/mL. The T-line on the nitrocellulose membrane was marked at density of 0.9 mL/cm. Goat anti-mouse H-FABP antibody (0.2 mg/mL) was immobilized on the C-line at a density of 0.9 mL/cm. After application of the T-line and C-line, the nitrocellulose membrane was dried at room temperature for 8 hours. Finally, 80-mL solutions of different H-FABP/gold colloid-conjugated antibody complex solutions were applied to test cards on which the T-line was prepared by immobilizing varying amounts of capture antibody: 0.5 mg/mL (a), 0.75 mg/mL (b), 1.0 mg/mL (c), and 1.5 mg/mL (d).

Article Snippet: Anti-human H-FABP antibody (FABP3 2302 SPTN-5, 5.1 mg/mL) and anti-human H-FABP antibody (FABP3 2304 SPTN-5, 5.0 mg/mL) were obtained from Medix Biochemica.

Techniques: Fluorescence, Concentration Assay, Membrane

Journal: Journal of Medical Biochemistry

Article Title: Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background Fluorescence Quenching Immune Chromatography

doi: 10.2478/jomb-2018-0017

Figure Lengend Snippet: Validation with H-FABP standard solutions: H-FABP concentration in solution vs. F 0 /F 1 .

Article Snippet: Anti-human H-FABP antibody (FABP3 2302 SPTN-5, 5.1 mg/mL) and anti-human H-FABP antibody (FABP3 2304 SPTN-5, 5.0 mg/mL) were obtained from Medix Biochemica.

Techniques: Biomarker Discovery, Concentration Assay

Journal: Journal of Medical Biochemistry

Article Title: Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background Fluorescence Quenching Immune Chromatography

doi: 10.2478/jomb-2018-0017

Figure Lengend Snippet: Repeatability of the test results.

Article Snippet: Anti-human H-FABP antibody (FABP3 2302 SPTN-5, 5.1 mg/mL) and anti-human H-FABP antibody (FABP3 2304 SPTN-5, 5.0 mg/mL) were obtained from Medix Biochemica.

Techniques: Concentration Assay

Journal: Journal of Medical Biochemistry

Article Title: Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background Fluorescence Quenching Immune Chromatography

doi: 10.2478/jomb-2018-0017

Figure Lengend Snippet: Test results for different sample batches.

Article Snippet: Anti-human H-FABP antibody (FABP3 2302 SPTN-5, 5.1 mg/mL) and anti-human H-FABP antibody (FABP3 2304 SPTN-5, 5.0 mg/mL) were obtained from Medix Biochemica.

Techniques: Concentration Assay

Journal: Journal of Medical Biochemistry

Article Title: Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background Fluorescence Quenching Immune Chromatography

doi: 10.2478/jomb-2018-0017

Figure Lengend Snippet: Potential interference of co-present substances in H-FABP solutions.

Article Snippet: Anti-human H-FABP antibody (FABP3 2302 SPTN-5, 5.1 mg/mL) and anti-human H-FABP antibody (FABP3 2304 SPTN-5, 5.0 mg/mL) were obtained from Medix Biochemica.

Techniques: Concentration Assay