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: PLoS ONE

Article Title: Quantitative Proteomic Analysis of Niemann-Pick Disease, Type C1 Cerebellum Identifies Protein Biomarkers and Provides Pathological Insight

doi: 10.1371/journal.pone.0047845

Figure Lengend Snippet: (A) Representative 2D-GE images of FABP3 showing the over-expression in the mutant mouse cerebellum compared to the control tissue at the week five time point (top) and graphically displayed (bottom). Arrows note the protein spot of interest. (B) FABP3 levels measured in CSF from control (n = 30) and NPC1 patients (n = 42). Data is represented as average ± standard error of the mean. Significance was determined using an unpaired t-test with Welch's correction, p<0.0001.

Article Snippet: The resulting cDNA was used for the following TaqMan Assays: Fabp3 (Mm02342494) and Gapdh (Rodent GAPDH Control Reagents).

Techniques: Over Expression, Mutagenesis, Control

Journal: PLoS ONE

Article Title: Quantitative Proteomic Analysis of Niemann-Pick Disease, Type C1 Cerebellum Identifies Protein Biomarkers and Provides Pathological Insight

doi: 10.1371/journal.pone.0047845

Figure Lengend Snippet: (A) Comparison of FABP3 concentration in untreated (n = 27) and miglustat treated NPC1 patients (n = 18) (p<0.01, t-test, unpaired, Welch's correction). (B) Serial change in FABP3 levels pre- and post-miglustat treatment. Lines connect measurements from the same patient before and after miglustat initiation where one patient was followed serially before miglustat treatment. (C) Percent change in CSF-FABP3 concentration pre- and post-miglustat treatment. (D) Percent change of FABP3 concentration over time in the untreated, treated and pre- post-groups. A one-way ANOVA was used to determined significance (p<0.0001) of the FABP3 concentration change following miglustat initiation.

Article Snippet: The resulting cDNA was used for the following TaqMan Assays: Fabp3 (Mm02342494) and Gapdh (Rodent GAPDH Control Reagents).

Techniques: Comparison, Concentration Assay

Journal: PLoS ONE

Article Title: BMS309403 Stimulates Glucose Uptake in Myotubes through Activation of AMP-Activated Protein Kinase

doi: 10.1371/journal.pone.0044570

Figure Lengend Snippet: Western blotting analysis of AMPK phosphorylation in C2C12 myobubes overexpressing FABP3 with or without BMS309403 treatment.

Article Snippet: The HIV-based lentiviral vectors and cDNA encoding full-length human FABP3 were purchased from Genecopoeia (Rockville, MD, U.S.). pCMV-3tag vector was purchased from Agilent technologies (Santa Clara, CA, U.S.).

Techniques: Western Blot, Phospho-proteomics

Journal: PLoS ONE

Article Title: BMS309403 Stimulates Glucose Uptake in Myotubes through Activation of AMP-Activated Protein Kinase

doi: 10.1371/journal.pone.0044570

Figure Lengend Snippet: (A) Microscopy photography (×100) of C2C12 cell lines stably expressing lentiviral HIVU6-EGFP or HIVU6-shRNA against mFABP3 (shFABP3). (B) Quantitative real-time PCR analysis of FABP3 gene expression in differentiated C2C12 stable myotubes. (C) Western blotting analysis of AMPK signaling pathway following BMS309403 treatment in the control and FABP3 knocked-down C2C12 myotubes. (D) Densitometric analysis of phospho-ACC and phospho-AMPKα in C2C12 stable myotubes of EGFP or shFABP3 treated with DMSO or 20 µM BMS309403. C2C12 stable myotubes of EGFP treated with DMSO was used as control. There was significant difference between DMSO and BMS group, but no significant difference between EGFP + BMS and shFABP3+ BMS group.

Article Snippet: The HIV-based lentiviral vectors and cDNA encoding full-length human FABP3 were purchased from Genecopoeia (Rockville, MD, U.S.). pCMV-3tag vector was purchased from Agilent technologies (Santa Clara, CA, U.S.).

Techniques: Microscopy, Stable Transfection, Expressing, shRNA, Real-time Polymerase Chain Reaction, Gene Expression, Western Blot, Control