Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: FABP3 was strongly expressed in PV+ GABAergic interneurons of the ACC. A, Left, RT-PCR analysis of Fabp3 in different brain areas. Right, qPCR analysis of Fabp3 in different brain areas. Error bar indicates mean ± SEM. n = 6 mice per group. **p < 0.01 versus WT ACC. B–F, Confocal images showing colocalization of FABP3 and a pyramidal neuronal marker (Ng), three classical GABAergic neuronal makers (PV, SOM, and CR), or GAD67 in the ACC. B, Most FABP3+ structures do not show immunoreactivity for Ng. C, Confocal images showing FABP3 and PV colocalization in the ACC. D, F, Most FABP+ structures do not show immunoreactivity for SOM (D) or CR (E). F, Confocal images showing FABP3 and GAD67 colocalization in the ACC (arrowheads). B, C (bottom), D, E (bottom right), F (right), Enlarged images of the boxed area in the merged image. Scale bars, 50 μm. Str, Striatum.

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Marker

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Colocalization of PV, SOM, and CR with FABP3 in the ACC a

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Fabp3 gene ablation in the ACC caused upregulation of GAD67 expression and GABA synthesis. A, Quantitative analysis of the number of PV−, SOM−, and CR+ neurons in the ACC. PV, n = 8 sections, 4 mice; SOM, n = 12 sections, 6 mice; CR, n = 12 sections, 6 mice per group. B, Representative immunoblots (left) and quantitative densitometry analysis (right) probed with GABA-related antibodies. GAD67 and β-actin, n = 6 mice per group; GAD65, VGAT, and GABAAR, n = 5 mice per group; gephyrin, n = 5 and 6 WT and Fabp3 KO mice, respectively. C, Quantitative analysis of Gad67 mRNA expression in the ACC from WT and Fabp3 KO mice. n = 4 mice per group. D, Quantitative analysis of the GABA concentrations in ACC extracts from WT and Fabp3 KO mice by ELISA. n = 6 mice per group. E–G, There were no differences in GAD67 expression or GABA synthesis in the mPFC or AM of Fabp3 KO mice. E, Representative immunoblots (left) and quantitative densitometry analysis (right) probed with GAD67 antibody. n = 4 mice per group. F, Quantitative analysis of the Gad67 mRNA expression. n = 4 mice per group. G, Quantitative analysis of the GABA concentrations in the mPFC and AM extracts by ELISA. mPFC, n = 6 mice per group; AM, n = 4 and 6 WT and Fabp3 KO mice, respectively. H, I, Quantitative analysis of the DA and 5-HT concentrations in ACC extracts by ELISA. 5-HT, n = 6 mice per group; DA, n = 6 and 5 WT and Fabp3 KO mice, respectively. A–I, **p < 0.01 versus WT mice. J, Left, Representative immunoblots probed with FABP3 antibody. Right, Quantitative analysis of Gad67 mRNA in Neuro-2a cells. n = 4 per group. **p < 0.01 versus control. mock, Mock control; OE, Fabp3 overexpression; NC, negative control; KD, Fabp3 knockdown. Error bar indicates mean ± SEM.

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression, Negative Control

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Enhancement of inhibitory synaptic plasticity in the ACC of Fabp3 KO mice. A, Representative traces of mEPSCs and mIPSCs in ACC layer II/III pyramidal neurons. B, Plots of the mEPSC and mIPSC frequencies in WT and Fabp3 KO mice. C, Plots of the mEPSC and mIPSC amplitudes in WT and Fabp3 KO mice. n = 53 cells for WT and 59 cells for Fabp3 KO from 5 mice for mEPSCs. n = 52 cells for WT and 60 cells for Fabp3 KO from 5 mice for mIPSCs. **p < 0.01 versus WT mice. Error bar indicates mean ± SEM.

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Extracellular glutamate (Glu) concentrations in the ACC in freely moving animals. A, Dialysate Glu concentrations at baseline (basal) and after depolarization stimulation (High-K+) (left). Glu AUC before (basal) and after (High-K+) depolarization stimulation (right). *p < 0.05 versus wild-type mice. B, Normalization of the dialysate Glu signals indicated that the response to depolarization stimulation was enhanced in Fabp3 KO mice (left). The normalized Glu AUC after depolarization stimulation (right). A, B, Error bar indicates mean ± SEM. *p < 0.05 versus WT mice. n = 6 WT mice; n = 5 Fabp3 KO mice. C, D, Representative immunoblots (left) and quantitative densitometry analysis (right) probed with various antibodies. *p < 0.05; **p < 0.01 versus WT mice. n = 6 mice per group. Error bar indicates mean ± SEM.

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques: Western Blot

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Decreased binding of MeCP2 and HDAC1 to the Gad67 promoter region in the ACC of Fabp3 KO mice. A, Representative immunoblots (top) and quantitative densitometry analysis (bottom) probed with various antibodies are shown. n = 6 mice per group. B, ChIP assay of ACC extracts from WT or Fabp3 KO mice via anti-MeCP2 and HDAC1. The precipitated DNA was analyzed by qPCR with primers amplifying the Gad67 promoter region. **p < 0.01 versus WT mice. n = 5 WT mice. n = 4 Fabp3 KO mice. C, DNA methylation profile in the Gad67 promoter region determined by bisulfite sequencing. Black circles represent methylated CpGs. Open circles represent unmethylated sites. Top numbers indicate global percentages of methylated cytosines. D, Quantitative analysis of the Gad67 mRNA expression in the ACC. WT, n = 6; KO, n = 5; KO + MET, n = 6 mice per group. **p < 0.01 versus WT mice. ##p < 0.01 versus KO mice. E, F, Quantitative analysis of the SAM concentration by ELISA. n = 4 per group. **p < 0.01 versus mock control. MET treatment (5.2 mmol/kg, s.c., twice per day for 6 d). mock, Mock control; OE, Fabp3 overexpression. Error bar indicates mean ± SEM.

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques: Binding Assay, Western Blot, DNA Methylation Assay, Methylation Sequencing, Methylation, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Over Expression

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Amelioration of behavioral abnormalities in Fabp3 KO mice by chronic treatment with MET. A, Schematic of the behavioral tests and MET treatment (5.2 mmol/kg, s.c., twice per day). During the behavioral testing period, mice received saline or MET once per day after testing. B, OFT. Graph represents the time spent in the center area over a 5 min period for WT and Fabp3 KO mice. n = 14–17 mice per group. C, HBT. An illustrative example of the travel pathway in the HBT using video tracking software (top). Graphs represent the time spent in the center area, total number of head-dips, head-dip duration, and latency to first head-dip for the 5 min testing session (bottom). n = 13–15 mice per group. **p < 0.01 versus saline-treated WT mice. ##p < 0.01 versus saline-treated KO mice. D, NORT. Differences in exploratory preference were assessed between groups in the training (left) or test (right) sessions. n = 11–15 mice per group. **p < 0.01 versus familiar group. Sal, Saline treatment. MET treatment (5.2 mmol/kg, s.c., twice per day for 6 d). Error bar indicates mean ± SEM.

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques: Software

Journal: The Journal of Neuroscience

Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region

doi: 10.1523/JNEUROSCI.1285-18.2018

Figure Lengend Snippet: Locomotor activities during behavioral tests a

Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).

Techniques:

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: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a UMAP plot displaying 14 identified cell types within the goat mammary glands. Cells are annotated and colored by type. Subsets of cell types including luminal, basal, fibroblast, immune, endothelial cell are labeled. b The percentages of −4W (red) and +1 W (blue) cells in each cell type are shown in a UMAP plot. c UMAP plots showing the expression of selected marker genes in four luminal subtypes. d Pseudotemporal trajectory analysis of scRNA-seq data of luminal secretory cells is shown in a UMAP plot. e Changes in the proportion of LumSecP and LumSec cells in luminal cell types were identified by scRNA-seq data at −4W and +1 W. n = 3 goats per group. f Representative images of tissue immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI at −4W and +1 W. Scale bars, 50 μm. g Bar plots exhibiting the percentage of ALDH1A3-positive cells in luminal cells (labeled by KRT18) in f . n = 15 sections per group. h Representative images of tissue immunofluorescence staining for FABP3 (red), KRT18 (green), and DAPI at −4W and +1 W. Scale bars, 50 μm. i Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in h . n = 20 sections per group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in e , g , i .

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Labeling, Expressing, Marker, Immunofluorescence, Staining

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a Relative proportions of LumHR cells in total luminal cells identified by scRNA-seq data at −4W and +1 W. n = 3 goats per group. b , c Bar plots exhibiting the percentage of PGR - and ESR1 -positive cells within luminal cells in scRNA-seq data. n = 3 goats per group. d Representative images of immunofluorescence staining for PR (red), KRT18 (green), and DAPI (blue). Scale bars, 50 μm. e Bar plots exhibiting the percentage of PR-positive cells in luminal cells (labeled by KRT18) in ( d ). n = 5 goats per group. f Representative images of immunofluorescence staining for ER (red), KRT18 (green), and DAPI (blue). Scale bars, 50 μm. g Bar plots exhibiting the percentage of ER-positive cells in luminal cells (labeled by KRT18) in ( f ). n = 5 goats. h Violin plot showing the specific expression of PRLR in LumHR cells by scRNA-seq. i Heatmap displaying the transcriptional level of indicated genes related to milk protein and luminal differentiation in the goat mammary organoids ( n = 3 biological replicates) treated with or without prolactin and in the mammary tissue at −4W (non-lactation) and +1 W (lactation). n = 3 goats for tissues. j Proportions of luminal cell types in goat mammary organoids incubated with or without prolactin predicted by CIBERSORTx deconvolution. n = 3 biological replicates. k Representative images of immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI in mammary organoids incubated with or without prolactin. Scale bars, 10 μm. l Bar plots exhibiting the percentage of ALDH1A3-positive cells in luminal cells (labeled by KRT18) in ( k ). n = 14 domes in the control group and n = 10 domes in the prolactin treated group. m Representative images of immunofluorescence staining for FABP3 (red), KRT18 (green) and DAPI in mammary organoids incubated with or without prolactin. Scale bars, 10 μm. n Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in ( m ). n = 6 domes in the control group and n = 7 domes in the prolactin-treated group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in ( a – c , e , g , j , l , n ).

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Immunofluorescence, Staining, Labeling, Expressing, Incubation, Control

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a Schematic illustration of targeted ablation of LumHR cells using the prlr -promoter to drive expression of DTA. b Experimental setup used in AAV intraductally injected mammary gland of ROSA-DTA +/ − mice under RR. c Whole-mount staining with carmine alum of mammary glands from ROSA-DTA +/ − mice (lactation day 2) intraductally injected with AAV-pPrlr-Cre or AAV-Control . Scale bars, 4 mm (top) and 500 μm (bottom). d , e Immunohistochemical staining and quantification of ER-positive luminal cells in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control. n = 3 mice per group. Scale bars, 50 μm. f , g Immunohistochemical staining and quantification of PR-positive luminal cells in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . n = 3 mice per group. Scale bars, 50 μm. h , i Immunohistochemical staining and quantification of β-casein-positive alveoli number per mm 2 in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . n = 3 mice per group. Scale bars, 50 μm. j Representative images of immunofluorescence staining for FABP3 (red), KRT18 (green), and DAPI in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . Scale bars, 50 μm. k Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in j . n = 3 mice per group. l Representative images of immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Contro l. Scale bars, 50 μm. m Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in ( l ). n = 3 mice per group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in d , g , i , l , m .

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Expressing, Injection, Staining, Control, Immunohistochemical staining, Immunofluorescence, Labeling

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