Journal: Journal of Biological Chemistry

Article Title: Acidic Fibroblast Growth Factor (FGF) Potentiates Glial-mediated Neurotoxicity by Activating FGFR2 IIIb Protein

doi: 10.1074/jbc.m111.270470

Figure Lengend Snippet: FIGURE 1. A, expression of mRNAs of the four FGF receptor genes (FGFR1–4) in various cells. The lanes are: 1, 100-bp ladder; 2, microglia; 3, astrocytes; 4, THP-1 cells; 5, U373 cells; and 6, SH-SY5Y cells. GAPDH loading controls are shown in the lower panel. Expression of mRNAs (B) and proteins (C) for the two spliced variants of FGFR1 and -2 (FGFR1 IIIb, FGFR1 IIIc, FGFR2 IIIb, FGFR2 IIIc), in glial cells are indicated. B, lane 1, 100-bp ladder; 2, microglia; 3, astrocytes; 4, THP-1 cells; and 5, U373 cells. GAPDH loading controls are shown in the lower panel. Equalization of sample loading was assessed independently using GAPDH as the housekeeping protein. Three independent experiments were performed and these were representatives. C, 1, microglia; 2, astrocytes; 3, THP-1 cells; and 4, U373 cells. Equalization of sample loading was assessed independently using -tubulin as the housekeeping protein. Three independent experiments were performed and these were representatives. D and E, quantitative results. Values are mean S.E., n 3. D, notice that FGFR2 is the major FGFR in glial cells, but SH-SY5Y cells express small amounts of all four FGFR genes. E, FGFR2 IIIb is the major receptor in all the glial cells examined.

Article Snippet: For protein expression of FGFR1 IIIb, FGFR1 IIIc, FGFR2 IIIb, and FGFR2 IIIc, the following antibodies were utilized: monoclonal anti-FGFR1 antibody (MAB658, R&D Systems, 1/2,000), monoclonal anti-FGFR2 (MAB6843, R&D Systems, 1/2,000) and, as the secondary antibody, anti-mouse IgG (A3682, Sigma, 1:3000).

Techniques: Expressing

Journal: Molecular Cancer

Article Title: Targeting the heparin-binding domain of fibroblast growth factor receptor 1 as a potential cancer therapy

doi: 10.1186/s12943-015-0391-4

Figure Lengend Snippet: Expression of FGFR1 in cells from breast and bone tissue. a , FGFR mRNA transcript levels in normal and cancer cells. b , Fold change in FGFR mRNA based on expression in cancers cells relative to normal cells. c , FGFR protein levels in normal versus cancer cells. Results are from triplicate experiments and the Western blot is representative of the triplicates. (* p < 0.05)

Article Snippet: The anti-FGFR1 antibody (clone # MAB765, R&D Systems) is one of the very few neutralizing antibodies against FGFR1 that have reached the market.

Techniques: Expressing, Western Blot

Journal: Molecular Cancer

Article Title: Targeting the heparin-binding domain of fibroblast growth factor receptor 1 as a potential cancer therapy

doi: 10.1186/s12943-015-0391-4

Figure Lengend Snippet: Specificity of IMB-R1 for FGFR1. a , Binding of IMB-R1 and MAB765 to FGFR1 isoforms (representative blot from triplicate experiments). b , Affinity of IMB-R1 for FGFR isoforms (results are from triplicate experiments). c , Binding of FGFR to heparin in the presence or absence of IMB-R1 (results are from triplicate experiments). d , Binding of FGF2 to FGFR in the presence or absence of IMB-R1 (results are from triplicate experiments). e , FGFR phosphorylation stimulated by FGF2 in the presence or absence of IMB-R1 including the fold change of FGFR phosphorylation as determined by a comparison of means

Article Snippet: The anti-FGFR1 antibody (clone # MAB765, R&D Systems) is one of the very few neutralizing antibodies against FGFR1 that have reached the market.

Techniques: Binding Assay, Phospho-proteomics, Comparison

Journal: Molecular Cancer

Article Title: Targeting the heparin-binding domain of fibroblast growth factor receptor 1 as a potential cancer therapy

doi: 10.1186/s12943-015-0391-4

Figure Lengend Snippet: Effects of FGFR1 inhibitors on cell growth. a , Fold change in cell number 48 h after treatment with IMB-R1. b , Temporal change in MG63 cell number following treatment with IMB-R1 (1:250). c , Cell number following FGF2 treatment (20 ng/ml for MG63, 5 ng/ml for MDAMB468 and T47D) for 48 h. Cells were pre-treatment with IMB-R1 for 1 h before FGF2 treatment. d , Cell number (MG63) following 48 h treatment with IMB-R1 or antigen-purified IMB-R1. e , Cell numbers following 48 h treatment with varying doses of FGFR inhibitors. f , Cell numbers following 48 h treatment with the FGFR1 antibody (MAB765) at varying doses. Unless otherwise stated, IMB-R1 was applied at a dilution of 1:250. All experiments were performed in triplicate

Article Snippet: The anti-FGFR1 antibody (clone # MAB765, R&D Systems) is one of the very few neutralizing antibodies against FGFR1 that have reached the market.

Techniques: Purification

Journal: Molecular Cancer

Article Title: Targeting the heparin-binding domain of fibroblast growth factor receptor 1 as a potential cancer therapy

doi: 10.1186/s12943-015-0391-4

Figure Lengend Snippet: The effect of IMB-R1 on gene expression. Cells were treated with IMB-R1 (1:250 dilution) for 48 h and RNA extracted for microarray analysis. a , The heatmaps: Red, up-regulation; Green, down-regulation. b , Top 10 affected cellular functions by IMB-R1 as determined by Ingenuity Pathway Analysis. c , The number of common genes affected across the different cells. d , The antioxidant genes significantly regulated by IMB-R1. e , The proposed signaling mechanisms disrupted by IMB-R1 during FGF2/FGFR1 dependent cell growth and survival in cancer cells

Article Snippet: The anti-FGFR1 antibody (clone # MAB765, R&D Systems) is one of the very few neutralizing antibodies against FGFR1 that have reached the market.

Techniques: Gene Expression, Microarray

Journal: Molecular Cancer

Article Title: Targeting the heparin-binding domain of fibroblast growth factor receptor 1 as a potential cancer therapy

doi: 10.1186/s12943-015-0391-4

Figure Lengend Snippet: IMB-R1 targets FGFR1 in multiple human cancers. a , IMB-R1 histochemical staining from a human cancer tissue array. Right panel , enlarged image of boxed area highlighting increased FGFR1 expression (detected by IMB-R1) in breast cancer tissues from twenty separate donors compared with adjacent healthy breast tissue. b , The intensity of FGFR1 expression (detected by IMB-R1) was scored and the average scores for the various cancer tissues compared with those from adjacent healthy tissues

Article Snippet: The anti-FGFR1 antibody (clone # MAB765, R&D Systems) is one of the very few neutralizing antibodies against FGFR1 that have reached the market.

Techniques: Staining, Expressing