fgfr4  (Thermo Fisher)


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  • 98
    Name:
    FGFR4 Recombinant Human Protein (without Catalytic Activity Domain), His Tag
    Description:
    Fibroblast growth factor receptor 4 (FGFR4), recombinant human protein is supplied as a lyophilized powder. It is suitable for use in analysis of protein structure, and performing cell based assays. It can also be used as an immunogen, as a protein standard, or in other research applications.Since this product is a truncated protein and does not contain intracellular catalytic activity domain, it is not suitable for use in enzyme activity studies..This recombinant protein was expressed from a DNA sequence encoding the extracellular domain (Met 1-Asp 369) of human FGFR4 (NP_002002.3) fused to a polyhistidine tag at the C-terminus.Activity: Measured by its ability to inhibit FGF1-dependent proliferation of BALB/c 3T3 mouse fibroblasts. The ED50 for this effect is typically 0.1-0.5 ng/ml.Formulation: Lyophilized in PBS, pH7.4, 5% Mannitol, 5% Trehalose, and 0.02% Tween-80.Reconstitution: Dissolve the protein in sterile double distilled water to a concentration of 0.2 mg/ml or lower. It is recommended that the protein be aliquoted and be used as soon as possible. Store aliquots under sterile conditions at -20°C. Avoid repeated freeze-thaw cycles.Expiration Date: Expires one year from date of receipt when stored as instructed.This protein is manufactured by SINO Biological.
    Catalog Number:
    10538H08H25
    Price:
    None
    Applications:
    Protein Biology, Assay Standard
    Size:
    5 x 5 µg
    Category:
    Proteins, Enzymes, & Peptides, Receptors & Cell Surface Markers, Growth Factor⁄Cytokine⁄Chemokine Receptors
    Score:
    85
    Buy from Supplier
    Name:
    FGFR4 Recombinant Human Protein
    Description:
    Fibroblast growth factor receptor 4 (FGFR4), recombinant human protein is supplied as a lyophilized powder. It is suitable for use in performing cell based assays. It can also be used as a protein standard, or in other research applications. Since this product is a truncated protein and does not contain intracellular catalytic activity domain, it is not suitable for use in enzyme activity studies..This recombinant protein was expressed from a DNA sequence encoding the extracellular domain (Met 1-Asp 369) of human FGFR4 precursor (NP_002002.3) fused to the Fc region of human IgG1 at the C-terminus.Activity: Measured by its ability to inhibit FGF1-dependent proliferation of BALB/c 3T3 mouse fibroblasts. The ED50 for this effect is typically 2-6 ng/ml.Formulation: Lyophilized in PBS, pH7.4, 5% Mannitol, 5% Trehalose, and 0.02% Tween-80.Reconstitution: Dissolve the protein in sterile double distilled water to a concentration of 0.2 mg/ml or lower. It is recommended that the protein be aliquoted and be used as soon as possible. Store aliquots under sterile conditions at -20°C. Avoid repeated freeze-thaw cycles. Expiration Date: Expires one year from date of receipt when stored as instructed.This protein is manufactured by SINO Biological.
    Catalog Number:
    10538H02H25
    Price:
    None
    Applications:
    Enzyme & Protein Activity Assays|Protein Assay Controls, Reference Standards & Accessories|Protein Assays and Analysis|Protein Biology, Assay Standard
    Size:
    5 x 5 µg
    Category:
    Proteins, Enzymes, & Peptides, Receptors & Cell Surface Markers, Growth Factor⁄Cytokine⁄Chemokine Receptors
    Score:
    85
    Buy from Supplier


    Structured Review

    Thermo Fisher fgfr4
    Effects of Glycine-to-Arginine substitution at position 388 of <t>FGFR4</t> on sorting of FGFR4 into internal MVB vesicles
    Fibroblast growth factor receptor 4 (FGFR4), recombinant human protein is supplied as a lyophilized powder. It is suitable for use in performing cell based assays. It can also be used as a protein standard, or in other research applications. Since this product is a truncated protein and does not contain intracellular catalytic activity domain, it is not suitable for use in enzyme activity studies..This recombinant protein was expressed from a DNA sequence encoding the extracellular domain (Met 1-Asp 369) of human FGFR4 precursor (NP_002002.3) fused to the Fc region of human IgG1 at the C-terminus.Activity: Measured by its ability to inhibit FGF1-dependent proliferation of BALB/c 3T3 mouse fibroblasts. The ED50 for this effect is typically 2-6 ng/ml.Formulation: Lyophilized in PBS, pH7.4, 5% Mannitol, 5% Trehalose, and 0.02% Tween-80.Reconstitution: Dissolve the protein in sterile double distilled water to a concentration of 0.2 mg/ml or lower. It is recommended that the protein be aliquoted and be used as soon as possible. Store aliquots under sterile conditions at -20°C. Avoid repeated freeze-thaw cycles. Expiration Date: Expires one year from date of receipt when stored as instructed.This protein is manufactured by SINO Biological.
    https://www.bioz.com/result/fgfr4/product/Thermo Fisher
    Average 98 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    fgfr4 - by Bioz Stars, 2019-10
    98/100 stars

    Images

    1) Product Images from "A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation"

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation

    Journal:

    doi: 10.1097/MPH.0000000000000506

    Effects of Glycine-to-Arginine substitution at position 388 of FGFR4 on sorting of FGFR4 into internal MVB vesicles
    Figure Legend Snippet: Effects of Glycine-to-Arginine substitution at position 388 of FGFR4 on sorting of FGFR4 into internal MVB vesicles

    Techniques Used:

    Effects of Glycine-to-Arginine substitution at position 388 of FGFR4 on receptor degradation
    Figure Legend Snippet: Effects of Glycine-to-Arginine substitution at position 388 of FGFR4 on receptor degradation

    Techniques Used:

    FGFR4 Polymorphism Distribution Among Cases and Controls
    Figure Legend Snippet: FGFR4 Polymorphism Distribution Among Cases and Controls

    Techniques Used:

    FGFR4 Genotype and Neuroblastoma Patient Outcomes and Prognostic Factors
    Figure Legend Snippet: FGFR4 Genotype and Neuroblastoma Patient Outcomes and Prognostic Factors

    Techniques Used:

    EGFR and FGFR4 degradation after ligand exposure
    Figure Legend Snippet: EGFR and FGFR4 degradation after ligand exposure

    Techniques Used:

    FGFR4 Polymorphism Distribution Among Cases and Controls
    Figure Legend Snippet: FGFR4 Polymorphism Distribution Among Cases and Controls

    Techniques Used:

    2) Product Images from "Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus"

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2018.01705

    GRP patterning is defective in fgfr4 CRISPR mutants. Most representative expression patterns of pre-somitic GRP markers coco, xnr1 , and gdf3 in stage 17 (A,C,E) and stage 19 (B,D,F) . GRPs of control and fgfr4 F0 CRISPR animals; ventral view of GRPs, anterior is to the top. Graphs show percentages of embryos with differential expression patterns of coco, xnr1 , and gdf3 . ** p
    Figure Legend Snippet: GRP patterning is defective in fgfr4 CRISPR mutants. Most representative expression patterns of pre-somitic GRP markers coco, xnr1 , and gdf3 in stage 17 (A,C,E) and stage 19 (B,D,F) . GRPs of control and fgfr4 F0 CRISPR animals; ventral view of GRPs, anterior is to the top. Graphs show percentages of embryos with differential expression patterns of coco, xnr1 , and gdf3 . ** p

    Techniques Used: CRISPR, Expressing

    The paraxial myogenic mesoderm is mispatterned in fgfr4 CRISPR embryos. (A–F, G–J ) Dorsal-vegetal views showing expression of an array of mesodermal markers in stage 10 (A–F) and stage 12 (G–J) embryos. Expression of paraxial mesoderm markers myf5 and myoD is perturbed in fgfr4 CRISPR embryos. (K, L) Xbra expression in embryos bisected through their dorsal midline at stages 10.5 and 12; red arrowheads point at xbra expression in the involuted mesoderm. Graphs show percentages of embryos with normal vs. abnormal expression of each marker; N = 32–40 embryos per Control or fgfr4 CRISPR; ** p
    Figure Legend Snippet: The paraxial myogenic mesoderm is mispatterned in fgfr4 CRISPR embryos. (A–F, G–J ) Dorsal-vegetal views showing expression of an array of mesodermal markers in stage 10 (A–F) and stage 12 (G–J) embryos. Expression of paraxial mesoderm markers myf5 and myoD is perturbed in fgfr4 CRISPR embryos. (K, L) Xbra expression in embryos bisected through their dorsal midline at stages 10.5 and 12; red arrowheads point at xbra expression in the involuted mesoderm. Graphs show percentages of embryos with normal vs. abnormal expression of each marker; N = 32–40 embryos per Control or fgfr4 CRISPR; ** p

    Techniques Used: CRISPR, Expressing, Marker

    Fgfr4 F0 CRISPR editing disrupts LR development. (A,B) fgfr4 CRISPR tadpoles display organ laterality defects. Ventral view of a stage 45 live tadpole with a cardiac L-loop ( B ; outlined) and inverse gut coiling ( B ; dashed line and red arrowhead). (C) Percentages of tadpoles with laterality defects (L-loops and inverse gut coiling) for three different CRISPRs; tadpoles with L-loops and inverse gut coils were scored; only tadpoles with inverse but otherwise intact gut coiling were considered; animals with completely uncoiled guts were scored as normal, as this phenotype occurs in the control population as well. Tadpoles with both cardiac and gut looping defects were only counted once in this analysis. (D) Pitx2c expression in the LPM of tailbud stage animals (stage 28); red arrowhead indicates absent expression. (E) Percentages of stage 28 animals with different pitx2c phenotypes; **** p
    Figure Legend Snippet: Fgfr4 F0 CRISPR editing disrupts LR development. (A,B) fgfr4 CRISPR tadpoles display organ laterality defects. Ventral view of a stage 45 live tadpole with a cardiac L-loop ( B ; outlined) and inverse gut coiling ( B ; dashed line and red arrowhead). (C) Percentages of tadpoles with laterality defects (L-loops and inverse gut coiling) for three different CRISPRs; tadpoles with L-loops and inverse gut coils were scored; only tadpoles with inverse but otherwise intact gut coiling were considered; animals with completely uncoiled guts were scored as normal, as this phenotype occurs in the control population as well. Tadpoles with both cardiac and gut looping defects were only counted once in this analysis. (D) Pitx2c expression in the LPM of tailbud stage animals (stage 28); red arrowhead indicates absent expression. (E) Percentages of stage 28 animals with different pitx2c phenotypes; **** p

    Techniques Used: CRISPR, Expressing

    GRP morphology and identity are altered in fgfr4 CRISPR embryos. (A–D) GRPs of fgfr4 CRISPR animals are morphologically distinct, as shown by phalloidin (actin) and anti-acetylated tubulin (cilia) stain; phenotypes ranging from mild (B) to severe (C,D) , depending on loss of small mesodermal ciliated cells. (E–H) Higher magnification of GRPs shows loss of ciliated GRP area in fgfr4 CRISPR embryos (G, H) . (I–K) The pre-somitic, myoD positive portion of the GRP (outlined) is drastically reduced in fgfr4 CRISPR embryos, even in embryos in which the overall GRP morphology is preserved (J) . (L) Quantification of total GRP area, defined morphologically by small, ciliated cells, is reduced in fgfr4 CRISPR embryos. (M) The myoD positive area of the GRP, normalized to total GRP area, is specifically reduced in fgfr4 CRISPR embryos. Scale bars in (A–D, I–K) = 40 μm, in (E–H) = 20 μm. ** p
    Figure Legend Snippet: GRP morphology and identity are altered in fgfr4 CRISPR embryos. (A–D) GRPs of fgfr4 CRISPR animals are morphologically distinct, as shown by phalloidin (actin) and anti-acetylated tubulin (cilia) stain; phenotypes ranging from mild (B) to severe (C,D) , depending on loss of small mesodermal ciliated cells. (E–H) Higher magnification of GRPs shows loss of ciliated GRP area in fgfr4 CRISPR embryos (G, H) . (I–K) The pre-somitic, myoD positive portion of the GRP (outlined) is drastically reduced in fgfr4 CRISPR embryos, even in embryos in which the overall GRP morphology is preserved (J) . (L) Quantification of total GRP area, defined morphologically by small, ciliated cells, is reduced in fgfr4 CRISPR embryos. (M) The myoD positive area of the GRP, normalized to total GRP area, is specifically reduced in fgfr4 CRISPR embryos. Scale bars in (A–D, I–K) = 40 μm, in (E–H) = 20 μm. ** p

    Techniques Used: CRISPR, Staining

    Fgfr4 expression during early X. tropicalis development, detected by in situ hybridization. (A,B) Whole stage 19 embryos; expression is detected in the head region (2: eyes), lateral plate mesoderm (3), anterior somites (1a) and posterior pre-somitic mesoderm (1b). (C,D) Transcripts were not detected in stage 16 and 19 GRPs (arrowheads). (E,F) Stage 10.5 embryos, whole ( E , dorsal view) or bisected through the dorsal midline (F) ; fgfr4 is broadly expressed in the ectoderm (4) and dorsal marginal zone (5). (G,H) Stage 12 embryos, whole ( G , dorso-vegetal view) or bisected (H) ; fgfr4 is absent from the marginal zone (5), but is expressed in the anterior migrating involuted mesoderm (6).
    Figure Legend Snippet: Fgfr4 expression during early X. tropicalis development, detected by in situ hybridization. (A,B) Whole stage 19 embryos; expression is detected in the head region (2: eyes), lateral plate mesoderm (3), anterior somites (1a) and posterior pre-somitic mesoderm (1b). (C,D) Transcripts were not detected in stage 16 and 19 GRPs (arrowheads). (E,F) Stage 10.5 embryos, whole ( E , dorsal view) or bisected through the dorsal midline (F) ; fgfr4 is broadly expressed in the ectoderm (4) and dorsal marginal zone (5). (G,H) Stage 12 embryos, whole ( G , dorso-vegetal view) or bisected (H) ; fgfr4 is absent from the marginal zone (5), but is expressed in the anterior migrating involuted mesoderm (6).

    Techniques Used: Expressing, In Situ Hybridization

    3) Product Images from "Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma"

    Article Title: Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-12-56

    (A) FGF19 mRNA expression in cell extracts from HuH7, HepG2, PLC/PRF/5, HLE, HLF, JHH1, JHH2, JHH5, JHH6, and JHH7 cell lines was examined using quantitative RT-PCR ( n = 8) . FGF19 was expressed in all cell lines. ( B ) FGF19 protein levels in the supernatant media from ten cell lines and normal hepatocytes (normal HC) were assayed by ELISA. FGF19 was detected in the supernatant of all of them. ( C ) Using anti-FGF19 monoclonal antibodies, diffuse positive staining was demonstrated in the cytoplasm of HuH7, HepG2, HLE, HLF, and JHH7 cells. The closed rectangles indicate the same cells stained Immunohistochemically using control antibody. ( D ) FGFR4 mRNA expression in cell extracts from HuH7, HepG2, PLC/PRF/5, HLE, HLF, JHH1, JHH2, JHH5, JHH6, and JHH7 cell lines was examined using quantitative RT-PCR ( n = 8). FGFR4 was expressed in all cell lines. ( E ) Using anti-FGFR4 monoclonal antibodies, positive staining was demonstrated in the cell membranes of all of the above HCC lines. (Original magnifications ( C ) × 400), ( E ) × 400)) The closed rectangles indicate the same cells stained Immunohistochemically using control antibody.
    Figure Legend Snippet: (A) FGF19 mRNA expression in cell extracts from HuH7, HepG2, PLC/PRF/5, HLE, HLF, JHH1, JHH2, JHH5, JHH6, and JHH7 cell lines was examined using quantitative RT-PCR ( n = 8) . FGF19 was expressed in all cell lines. ( B ) FGF19 protein levels in the supernatant media from ten cell lines and normal hepatocytes (normal HC) were assayed by ELISA. FGF19 was detected in the supernatant of all of them. ( C ) Using anti-FGF19 monoclonal antibodies, diffuse positive staining was demonstrated in the cytoplasm of HuH7, HepG2, HLE, HLF, and JHH7 cells. The closed rectangles indicate the same cells stained Immunohistochemically using control antibody. ( D ) FGFR4 mRNA expression in cell extracts from HuH7, HepG2, PLC/PRF/5, HLE, HLF, JHH1, JHH2, JHH5, JHH6, and JHH7 cell lines was examined using quantitative RT-PCR ( n = 8). FGFR4 was expressed in all cell lines. ( E ) Using anti-FGFR4 monoclonal antibodies, positive staining was demonstrated in the cell membranes of all of the above HCC lines. (Original magnifications ( C ) × 400), ( E ) × 400)) The closed rectangles indicate the same cells stained Immunohistochemically using control antibody.

    Techniques Used: Expressing, Planar Chromatography, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining

    Real-time quantitative RT-PCR analysis, immunohistochemical staining of representative specimens from HCC patients of 40 HCC samples . ( A ): Ratio of average FGF19/GAPDH expression in HCC (T) compared with corresponding noncancerous hepatic tissues (N). The average FGF19/GAPDH level in HCCs. ( B ): The average FGFR4/GAPDH level in HCCs. ( C ): Immunohistochemistry using anti-FGF19 monoclonal antibodies; HCC tissue (lower) and noncancerous hepatocytes (upper). ( D ): Immunohistochemistry using anti-FGFR4 monoclonal antibodies; HCC tissue (lower) and noncancerous tissue (upper). (Original magnifications: ×40 (upper); ×40 (lower)). RT-PCR; reverse transcription polymerase chain reaction; HCC, hepatocellular carcinoma.
    Figure Legend Snippet: Real-time quantitative RT-PCR analysis, immunohistochemical staining of representative specimens from HCC patients of 40 HCC samples . ( A ): Ratio of average FGF19/GAPDH expression in HCC (T) compared with corresponding noncancerous hepatic tissues (N). The average FGF19/GAPDH level in HCCs. ( B ): The average FGFR4/GAPDH level in HCCs. ( C ): Immunohistochemistry using anti-FGF19 monoclonal antibodies; HCC tissue (lower) and noncancerous hepatocytes (upper). ( D ): Immunohistochemistry using anti-FGFR4 monoclonal antibodies; HCC tissue (lower) and noncancerous tissue (upper). (Original magnifications: ×40 (upper); ×40 (lower)). RT-PCR; reverse transcription polymerase chain reaction; HCC, hepatocellular carcinoma.

    Techniques Used: Quantitative RT-PCR, Immunohistochemistry, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

    4) Product Images from "Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus"

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2018.01705

    Fgfr4 F0 CRISPR editing disrupts LR development. (A,B) fgfr4 CRISPR tadpoles display organ laterality defects. Ventral view of a stage 45 live tadpole with a cardiac L-loop ( B ; outlined) and inverse gut coiling ( B ; dashed line and red arrowhead). (C) Percentages of tadpoles with laterality defects (L-loops and inverse gut coiling) for three different CRISPRs; tadpoles with L-loops and inverse gut coils were scored; only tadpoles with inverse but otherwise intact gut coiling were considered; animals with completely uncoiled guts were scored as normal, as this phenotype occurs in the control population as well. Tadpoles with both cardiac and gut looping defects were only counted once in this analysis. (D) Pitx2c expression in the LPM of tailbud stage animals (stage 28); red arrowhead indicates absent expression. (E) Percentages of stage 28 animals with different pitx2c phenotypes; **** p < 0.0001, *** p < 0.001.
    Figure Legend Snippet: Fgfr4 F0 CRISPR editing disrupts LR development. (A,B) fgfr4 CRISPR tadpoles display organ laterality defects. Ventral view of a stage 45 live tadpole with a cardiac L-loop ( B ; outlined) and inverse gut coiling ( B ; dashed line and red arrowhead). (C) Percentages of tadpoles with laterality defects (L-loops and inverse gut coiling) for three different CRISPRs; tadpoles with L-loops and inverse gut coils were scored; only tadpoles with inverse but otherwise intact gut coiling were considered; animals with completely uncoiled guts were scored as normal, as this phenotype occurs in the control population as well. Tadpoles with both cardiac and gut looping defects were only counted once in this analysis. (D) Pitx2c expression in the LPM of tailbud stage animals (stage 28); red arrowhead indicates absent expression. (E) Percentages of stage 28 animals with different pitx2c phenotypes; **** p < 0.0001, *** p < 0.001.

    Techniques Used: CRISPR, Expressing

    5) Product Images from "Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition"

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition

    Journal:

    doi: 10.1002/ijc.29893

    FGFR4 knockdown suppresses S2 HCC cell proliferation
    Figure Legend Snippet: FGFR4 knockdown suppresses S2 HCC cell proliferation

    Techniques Used:

    6) Product Images from "Differential Effects of Polymorphic Alleles of FGF Receptor 4 on Colon Cancer Growth and Metastasis"

    Article Title: Differential Effects of Polymorphic Alleles of FGF Receptor 4 on Colon Cancer Growth and Metastasis

    Journal:

    doi: 10.1158/0008-5472.CAN-11-3654

    Impact of FGFR4 overexpression on tumor cell growth and migration in vitro . Stable transfectants overexpressing FGFR4arg or FGFR4gly in SW480, HCT116, or HT29 cells were used to assess growth and malignant characteristics in vitro . Stable transfectants with the pcDNA3 vector were used as controls. A, 100 and 200 cells were seeded/6-well, medium was changed after 24 hours, and the number of colonies counted after 10 days of growth. B, five thousand cells each were suspended in soft agar medium and plated in 6-well plates. The number of colonies was counted at a magnification of 10-fold after 3 weeks of growth. C, cell migration was determined by filter migration assay from 2 × 104 cells/24-well. Photographs of cultures show representative results from SW480 transfectants. Quantitative results from all cell lines were calculated as % of control, pooled from at least 3 independent experiments, and presented as mean ± SD. *, **, *** indicate an increase as compared with the control at P < 0.05, 0.01, and 0.001, respectively. ## indicates a decrease as compared with control at P < 0.01. & , & & , and & & & indicate a difference between the FGFR4gly and FGFR4arg groups at P < 0.05, 0.01, and 0.001, respectively.
    Figure Legend Snippet: Impact of FGFR4 overexpression on tumor cell growth and migration in vitro . Stable transfectants overexpressing FGFR4arg or FGFR4gly in SW480, HCT116, or HT29 cells were used to assess growth and malignant characteristics in vitro . Stable transfectants with the pcDNA3 vector were used as controls. A, 100 and 200 cells were seeded/6-well, medium was changed after 24 hours, and the number of colonies counted after 10 days of growth. B, five thousand cells each were suspended in soft agar medium and plated in 6-well plates. The number of colonies was counted at a magnification of 10-fold after 3 weeks of growth. C, cell migration was determined by filter migration assay from 2 × 104 cells/24-well. Photographs of cultures show representative results from SW480 transfectants. Quantitative results from all cell lines were calculated as % of control, pooled from at least 3 independent experiments, and presented as mean ± SD. *, **, *** indicate an increase as compared with the control at P < 0.05, 0.01, and 0.001, respectively. ## indicates a decrease as compared with control at P < 0.01. & , & & , and & & & indicate a difference between the FGFR4gly and FGFR4arg groups at P < 0.05, 0.01, and 0.001, respectively.

    Techniques Used: Over Expression, Migration, In Vitro, Plasmid Preparation

    Related Articles

    Clone Assay:

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    Article Snippet: His-tagged human FAK-CD (AA 677-1052) was cloned into pET15b vector and similarly purified on Ni-NTA resin (Thermo Scientific). .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    Amplification:

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    Article Snippet: Paragraph title: Quantitative real time RT-PCR amplification of cDNAs ... The probes and primer kits used for each gene were as follows: Hs00172113-m1 for VDR, Hs00168017-m1 for CYP27B1, Hs00167999-m1 for CYP24A1, Hs00189038-m1 for LL37, Hs00915134-g1 for FGFR1, Hs01552926-m1 for FGFR2, Hs00179829-m1 for FGFR3, Hs01106908-m1 for FGFR4, Hs00183100-m1 for Klotho and Hs00174128-m1 for TNFα (Applied Biosystems).

    Construct:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
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    Real-time Polymerase Chain Reaction:

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select). .. Lipofectamine RNAiMax Reagent and 10 μM stock siRNA were dissolved in a 1:1 ratio in Opti-MEM Media, incubated for 5 minutes, and added to wells to achieve a final concentration of 30 nM siRNA.

    Incubation:

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation
    Article Snippet: After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000). .. After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000).

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs. .. Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs.

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs. .. Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs.

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select). .. Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select).

    Expressing:

    Article Title: Differential Effects of Polymorphic Alleles of FGF Receptor 4 on Colon Cancer Growth and Metastasis
    Article Snippet: Paragraph title: Knockdown of gene expression ... siRNAs specifically targeting FGFR4 were purchased from Ambion (Applied Biosystems) and transfected into 70% confluent cultures kept in medium containing 10% FCS using 3 μL siLentFect (BioRad) and 20 pmol of the siRNA per well in culture medium without serum.

    Article Title: FGF23 inhibits extra-renal synthesis of 1,25-dihydroxyvitamin D in human monocytes
    Article Snippet: PCR amplification of target gene cDNA was conducted using Taqman human gene expression assays, as previously described ( ). .. The probes and primer kits used for each gene were as follows: Hs00172113-m1 for VDR, Hs00168017-m1 for CYP27B1, Hs00167999-m1 for CYP24A1, Hs00189038-m1 for LL37, Hs00915134-g1 for FGFR1, Hs01552926-m1 for FGFR2, Hs00179829-m1 for FGFR3, Hs01106908-m1 for FGFR4, Hs00183100-m1 for Klotho and Hs00174128-m1 for TNFα (Applied Biosystems).

    Modification:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: His-tagged avian FAK-FERM (AA 31-405) in modified pET vector (provided by Dr. Michael Eck) was expressed in BL21 (DE3) E. coli (Life Technologies) and purified on Ni-NTA resin (Thermo Scientific) using buffers as described ( ). .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    Western Blot:

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation
    Article Snippet: Control reactions and experimental reaction pellets were resuspended in sample buffer. .. After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000). .. Proteins were detected with enhanced chemiluminescence (ECL; Pierce) and exposed to X-ray film.

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select). .. Lipofectamine RNAiMax Reagent and 10 μM stock siRNA were dissolved in a 1:1 ratio in Opti-MEM Media, incubated for 5 minutes, and added to wells to achieve a final concentration of 30 nM siRNA.

    Transfection:

    Article Title: Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma
    Article Snippet: We examined migration assay with 6 chambers in the same condition at the same time. .. Transfections of FGF19, FGFR4 and non-targeting negative control small interfering RNA (siRNA) (AMBION, Austin, TX) were conducted with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) in 96-well plates according to the manufacturer's specifications. .. The day before transfection, the JHH7 cells were trypsinized, counted, and seeded at 6 × 103 cells per well into 96-well plates.

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: Paragraph title: Transfection of short interfering ribonucleic acid (siRNA) ... Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select).

    Article Title: Differential Effects of Polymorphic Alleles of FGF Receptor 4 on Colon Cancer Growth and Metastasis
    Article Snippet: Controls were transfected with GFP or the pcDNA3 vector and stable transfectants were selected in the presence of geneticin (G418). .. siRNAs specifically targeting FGFR4 were purchased from Ambion (Applied Biosystems) and transfected into 70% confluent cultures kept in medium containing 10% FCS using 3 μL siLentFect (BioRad) and 20 pmol of the siRNA per well in culture medium without serum. .. A scrambled siRNA without sequence homology to known human genes served as negative control.

    In Situ Hybridization:

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: Paragraph title: In situ Hybridization ... Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs.

    Sequencing:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: GST-HER2-ICD (AA 676-1255), GST-HER2-ICD1 (AA 676-801), GST-HER2-ICD2 (AA 802-1029), and GST-HER2-ICD3 (AA 1030-1255) constructs were designed into the pGEX-4T1 vector, sequenced by the Roswell Park Sequencing Core, expressed in BL21 (DE3) E. coli, and purified on Glutathione Sepharose 4B resin (GE Healthcare). .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    Recombinant:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: HER2-ECD protein (cat#BMS362) was purchased from eBiocience. .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies. .. Purified FAK-FERM (100ng) or FAK-CD (100ng) were incubated with purified HER2 (100ng) or additional RTK for 30 min in the presence or absence of ATP (10µM) in standard tyrosine kinase buffer: 20mM HEPES (pH=7.3), 5mM MgCl2 , 5mM MnCl2 , 2mM DTT, 0.1mg/mL BSA, and 0.1mM Na3 VO4 .

    Isolation:

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation
    Article Snippet: For reactions with added cytosol, 15μL endosomal membranes were mixed with 6μL ATP regeneration system (2mM MgATP, 50μg/mL creatine kinase, 8mM phosphocreatine, 1mM DTT), 25μg mammalian cytosol (isolated as above) and homogenization buffer to a final volume of 50μL. .. After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000).

    Article Title: Differential Effects of Polymorphic Alleles of FGF Receptor 4 on Colon Cancer Growth and Metastasis
    Article Snippet: siRNAs specifically targeting FGFR4 were purchased from Ambion (Applied Biosystems) and transfected into 70% confluent cultures kept in medium containing 10% FCS using 3 μL siLentFect (BioRad) and 20 pmol of the siRNA per well in culture medium without serum. .. A scrambled siRNA without sequence homology to known human genes served as negative control.

    Negative Control:

    Article Title: Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma
    Article Snippet: We examined migration assay with 6 chambers in the same condition at the same time. .. Transfections of FGF19, FGFR4 and non-targeting negative control small interfering RNA (siRNA) (AMBION, Austin, TX) were conducted with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) in 96-well plates according to the manufacturer's specifications. .. The day before transfection, the JHH7 cells were trypsinized, counted, and seeded at 6 × 103 cells per well into 96-well plates.

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: IC50 value was defined as the drug concentration yielding 50% nonsurviving cells compared with vehicle-treated controls. .. Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select). .. Validated siRNA against FGFR2 (cat#S102665299) was obtained from Qiagen (FlexiTube). siRNA transfection was done according to the protocol supplied by Life Technologies.

    Labeling:

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs. .. Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs.

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs. .. Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs.

    Purification:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: GST-HER2-ICD (AA 676-1255), GST-HER2-ICD1 (AA 676-801), GST-HER2-ICD2 (AA 802-1029), and GST-HER2-ICD3 (AA 1030-1255) constructs were designed into the pGEX-4T1 vector, sequenced by the Roswell Park Sequencing Core, expressed in BL21 (DE3) E. coli, and purified on Glutathione Sepharose 4B resin (GE Healthcare). .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    Protein Purification:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: Paragraph title: Protein Purification ... Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    Polymerase Chain Reaction:

    Article Title: FGF23 inhibits extra-renal synthesis of 1,25-dihydroxyvitamin D in human monocytes
    Article Snippet: PCR amplification of target gene cDNA was conducted using Taqman human gene expression assays, as previously described ( ). .. The probes and primer kits used for each gene were as follows: Hs00172113-m1 for VDR, Hs00168017-m1 for CYP27B1, Hs00167999-m1 for CYP24A1, Hs00189038-m1 for LL37, Hs00915134-g1 for FGFR1, Hs01552926-m1 for FGFR2, Hs00179829-m1 for FGFR3, Hs01106908-m1 for FGFR4, Hs00183100-m1 for Klotho and Hs00174128-m1 for TNFα (Applied Biosystems).

    Small Interfering RNA:

    Article Title: Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma
    Article Snippet: We examined migration assay with 6 chambers in the same condition at the same time. .. Transfections of FGF19, FGFR4 and non-targeting negative control small interfering RNA (siRNA) (AMBION, Austin, TX) were conducted with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) in 96-well plates according to the manufacturer's specifications. .. The day before transfection, the JHH7 cells were trypsinized, counted, and seeded at 6 × 103 cells per well into 96-well plates.

    Quantitative RT-PCR:

    Article Title: FGF23 inhibits extra-renal synthesis of 1,25-dihydroxyvitamin D in human monocytes
    Article Snippet: Paragraph title: Quantitative real time RT-PCR amplification of cDNAs ... The probes and primer kits used for each gene were as follows: Hs00172113-m1 for VDR, Hs00168017-m1 for CYP27B1, Hs00167999-m1 for CYP24A1, Hs00189038-m1 for LL37, Hs00915134-g1 for FGFR1, Hs01552926-m1 for FGFR2, Hs00179829-m1 for FGFR3, Hs01106908-m1 for FGFR4, Hs00183100-m1 for Klotho and Hs00174128-m1 for TNFα (Applied Biosystems).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: IC50 value was defined as the drug concentration yielding 50% nonsurviving cells compared with vehicle-treated controls. .. Scrambled negative control siRNA (cat4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select). .. Validated siRNA against FGFR2 (cat#S102665299) was obtained from Qiagen (FlexiTube). siRNA transfection was done according to the protocol supplied by Life Technologies.

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: HER2-ECD protein (cat#BMS362) was purchased from eBiocience. .. Recombinant HER2-ICD (catPV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies. .. Purified FAK-FERM (100ng) or FAK-CD (100ng) were incubated with purified HER2 (100ng) or additional RTK for 30 min in the presence or absence of ATP (10µM) in standard tyrosine kinase buffer: 20mM HEPES (pH=7.3), 5mM MgCl2 , 5mM MnCl2 , 2mM DTT, 0.1mg/mL BSA, and 0.1mM Na3 VO4 .

    SDS Page:

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation
    Article Snippet: Control reactions and experimental reaction pellets were resuspended in sample buffer. .. After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000). .. Proteins were detected with enhanced chemiluminescence (ECL; Pierce) and exposed to X-ray film.

    Plasmid Preparation:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: GST-HER2-ICD (AA 676-1255), GST-HER2-ICD1 (AA 676-801), GST-HER2-ICD2 (AA 802-1029), and GST-HER2-ICD3 (AA 1030-1255) constructs were designed into the pGEX-4T1 vector, sequenced by the Roswell Park Sequencing Core, expressed in BL21 (DE3) E. coli, and purified on Glutathione Sepharose 4B resin (GE Healthcare). .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    Multiplex Assay:

    Article Title: FGF23 inhibits extra-renal synthesis of 1,25-dihydroxyvitamin D in human monocytes
    Article Snippet: All reactions were normalized by multiplex analysis with the housekeeping 18S rRNA gene (Applied Biosystems, Foster City, CA, USA). .. The probes and primer kits used for each gene were as follows: Hs00172113-m1 for VDR, Hs00168017-m1 for CYP27B1, Hs00167999-m1 for CYP24A1, Hs00189038-m1 for LL37, Hs00915134-g1 for FGFR1, Hs01552926-m1 for FGFR2, Hs00179829-m1 for FGFR3, Hs01106908-m1 for FGFR4, Hs00183100-m1 for Klotho and Hs00174128-m1 for TNFα (Applied Biosystems).

    Positron Emission Tomography:

    Article Title: Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-kinase Inhibitors
    Article Snippet: His-tagged avian FAK-FERM (AA 31-405) in modified pET vector (provided by Dr. Michael Eck) was expressed in BL21 (DE3) E. coli (Life Technologies) and purified on Ni-NTA resin (Thermo Scientific) using buffers as described ( ). .. Recombinant HER2-ICD (cat#PV3366), EGFR (cat#PV3872), Tie2 (cat#PV3628), EphA2 (cat#PV3688), and FGFR4 (cat#P3054) were purchased from Life Technologies.

    In Vitro:

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: D-loops were defined as outflow tracts directed to the right, and L-loops to the left. .. Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs. .. Embryos were collected at the desired stages, fixed in MEMFA for 1–2 h at room temperature (RT) and dehydrated in 100% ethanol.

    Article Title: Candidate Heterotaxy Gene FGFR4 Is Essential for Patterning of the Left-Right Organizer in Xenopus
    Article Snippet: D-loops were defined as outflow tracts directed to the right, and L-loops to the left. .. Digoxigenin-labeled antisense probes for pitx2 (TNeu083k20), coco (TEgg007d24), xnr1 (TGas124h10), gdf3 (Tgas137g21), gsc (TNeu077f20), xnr3 (Tgas011k18), foxj1 (Tneu058M03), vent2 (BG885317), myf5 (TGas127b01), xbra (TNeu024F07), fgfr4 (Dharmacon, Clone ID: 7521919) were in vitro transcribed using T7 High Yield RNA Synthesis Kit (E2040S) from New England Biolabs. .. Embryos were collected at the desired stages, fixed in MEMFA for 1–2 h at room temperature (RT) and dehydrated in 100% ethanol.

    Tube Formation Assay:

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation
    Article Snippet: Paragraph title: Cell-free reconstitution of multivesicular body formation assay ... After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000).

    Homogenization:

    Article Title: A Polymorphism in the FGFR4 Gene is associated with Risk of Neuroblastoma and Altered Receptor Degradation
    Article Snippet: For reactions with added cytosol, 15μL endosomal membranes were mixed with 6μL ATP regeneration system (2mM MgATP, 50μg/mL creatine kinase, 8mM phosphocreatine, 1mM DTT), 25μg mammalian cytosol (isolated as above) and homogenization buffer to a final volume of 50μL. .. After boiling, proteins were separated by SDS-PAGE, followed by Western blotting with a murine antibody against the intracellular V5 tag of FGFR4 (Invitrogen, 1:5000) followed by goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP) (Sigma-Aldrich Inc, 1:10,000).

    Functional Assay:

    Article Title: Differential Effects of Polymorphic Alleles of FGF Receptor 4 on Colon Cancer Growth and Metastasis
    Article Snippet: siRNAs specifically targeting FGFR4 were purchased from Ambion (Applied Biosystems) and transfected into 70% confluent cultures kept in medium containing 10% FCS using 3 μL siLentFect (BioRad) and 20 pmol of the siRNA per well in culture medium without serum. .. After 24 and 48 hours, RNA and protein were isolated to verify knockdown efficiency.

    Concentration Assay:

    Article Title: Fibroblast growth factor 19 expression correlates with tumor progression and poorer prognosis of hepatocellular carcinoma
    Article Snippet: Transfections of FGF19, FGFR4 and non-targeting negative control small interfering RNA (siRNA) (AMBION, Austin, TX) were conducted with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) in 96-well plates according to the manufacturer's specifications. .. Transfections of FGF19, FGFR4 and non-targeting negative control small interfering RNA (siRNA) (AMBION, Austin, TX) were conducted with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) in 96-well plates according to the manufacturer's specifications.

    Article Title: Molecular Subclasses of Hepatocellular Carcinoma Predict Sensitivity to Fibroblast Growth Factor Receptor Inhibition
    Article Snippet: Scrambled negative control siRNA (cat#4390843) and validated siRNA against FGFR1 (id-s5164), FGFR3 (id-s5168), and FGFR4 (id S5177 and 1412) were all obtained from Life Technologies (Silencer Select). .. 1 × 105 cells were seeded into six-well plates, incubated overnight, then washed in cold PBS and changed to antibiotic free media immediately prior to transfection.

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    Thermo Fisher gene exp fgfr4 hs00242558 m1
    Mutations promote <t>FGFR4</t> autophosphorylation, Stat3 phosphorylation, and activation of cell cycle and DNA replication pathways.
    Gene Exp Fgfr4 Hs00242558 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 907 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp fgfr4 hs00242558 m1/product/Thermo Fisher
    Average 85 stars, based on 907 article reviews
    Price from $9.99 to $1999.99
    gene exp fgfr4 hs00242558 m1 - by Bioz Stars, 2019-10
    85/100 stars
      Buy from Supplier

    97
    Thermo Fisher gene exp fgfr4 hs01106908 m1
    N-cadherin is involved in the pro-oncogenic role of Arg388 <t>FGFR4</t> overexpression. ( A ) N-cadherin silencing in the FGFR4-388Arg-overexpressing H2009 cell line using a shRNA approach. Clonability ( B ) and soft agar assays ( C ) of the FGFR4-388Arg-overexpressing, N-cadherin silenced H2009 cell line. ( D ) Relative tumor growth of the xenograft in immunodeprived nude mice of these cell lines. ( E ) Western blot showing the STAT3 activation suspension by the STAT3 inhibitor SI3-201 along the time, which is accompanied by N-cadherin protein levels reduction, in the FGFR4-388Arg-overexpressing H2009 and H226 cell lines. SI3-201 exposure in hours is indicated in the figure. ( F ) Western blot showing the effect of STAT3 inhibition and N-cadherin shRNA silencing in several cancer-related downstream signaling pathways in the EV, FGFR4-388Gly and -388Arg-overexpressing H2009 cell line. In soft agar and clonability assays, colony number representation is shown. All values were normalized to empty vector control for each replicate of the experiment and the mean and standard deviation of every normalized replicate are represented. For western blots, a representative blot is shown. EV = Empty Vector control, FGFR4-Gly = FGFR4-388Gly overexpressing, FGFR4-Arg = FGFR4-388Arg overexpressing. p-Values are represented as asterisks (*p
    Gene Exp Fgfr4 Hs01106908 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp fgfr4 hs01106908 m1/product/Thermo Fisher
    Average 97 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    gene exp fgfr4 hs01106908 m1 - by Bioz Stars, 2019-10
    97/100 stars
      Buy from Supplier

    93
    Thermo Fisher gene exp fgfr4 mm01341852 m1
    <t>FGFR4</t> 80-bp deletion line lost FGFR4 expression and signaling. A , genotyping characterization of the 80-bp deletion FGFR4 KO animals. A predicted 279-bp band in WT littermate control animals, 199 bp in KO animals, and both bands in heterozygous ( HET )
    Gene Exp Fgfr4 Mm01341852 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp fgfr4 mm01341852 m1/product/Thermo Fisher
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    gene exp fgfr4 mm01341852 m1 - by Bioz Stars, 2019-10
    93/100 stars
      Buy from Supplier

    Image Search Results


    Mutations promote FGFR4 autophosphorylation, Stat3 phosphorylation, and activation of cell cycle and DNA replication pathways.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: Mutations promote FGFR4 autophosphorylation, Stat3 phosphorylation, and activation of cell cycle and DNA replication pathways.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: Activation Assay

    FGFR4 suppression leads to inhibition of in vivo growth and lung metastasis.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 suppression leads to inhibition of in vivo growth and lung metastasis.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: Inhibition, In Vivo

    FGFR4 mutations transform 3T3 cells.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 mutations transform 3T3 cells.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques:

    FGFR4 mutations accelerate growth and promote a metastasis phenotype.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 mutations accelerate growth and promote a metastasis phenotype.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques:

    FGFR4 expression in RMS shows correlation with FGFR4 protein, advanced stage, ARMS histology, and poor survival.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 expression in RMS shows correlation with FGFR4 protein, advanced stage, ARMS histology, and poor survival.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: Expressing

    Oncogene dependence and inhibition of FGFR4 .

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: Oncogene dependence and inhibition of FGFR4 .

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: Inhibition

    FGFR4 knockdown with an inducible shRNA leads to reduced in vitro growth.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 knockdown with an inducible shRNA leads to reduced in vitro growth.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: shRNA, In Vitro

    Identification of FGFR4 TK domain mutations in human RMS tumors.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: Identification of FGFR4 TK domain mutations in human RMS tumors.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques:

    Structural modeling of the FGFR4 codon 535 and 550 mutations.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: Structural modeling of the FGFR4 codon 535 and 550 mutations.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques:

    FGFR4 TK domain mutations in RMS.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 TK domain mutations in RMS.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques:

    FGFR4 mutations promote FGFR4 autophosphorylation, STAT3 phosphorylation, and activation of cell cycle and DNA replication pathways.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: FGFR4 mutations promote FGFR4 autophosphorylation, STAT3 phosphorylation, and activation of cell cycle and DNA replication pathways.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: Activation Assay

    High FGFR4 expression in RMS is associated with advanced stage, ARMS histology, and poor survival.

    Journal:

    Article Title: Identification of FGFR4-activating mutations in human rhabdomyosarcomas that promote metastasis in xenotransplanted models

    doi: 10.1172/JCI39703

    Figure Lengend Snippet: High FGFR4 expression in RMS is associated with advanced stage, ARMS histology, and poor survival.

    Article Snippet: Assays-on-Demand (ABI) were used for assessing FGFR4 expression levels (primer Hs00242558_m1), and fold change was determined by normalizing to GAPDH (Hs99999905_m1).

    Techniques: Expressing

    In vivo tumor growth assay with daily treatment of 30 mg/kg of ponatinib after tumor volumes reach 100 mm 3 . Arrow indicates the start of treatment. (A) Treatment of tumors harboring the FGFR4 N535K mutation with ponatinib significantly inhibits tumor growth after 10 days of treatment (*p = 0.0165, **p = 0.0048). (B) Treatment of tumors containing the FGFR4 V550E mutation with ponatinib significantly inhibits tumor growth after 6 days of treatment (*p = 0.0185, **p = 0.0087, ***p = 0.0005). (C) Treatment of tumors expressing the wild-type FGFR4 with ponatinib does not affect tumor growth. (D) Treatment of tumors expressing the empty vector with ponatinib does not affect tumor growth.

    Journal: PLoS ONE

    Article Title: Targeting Wild-Type and Mutationally Activated FGFR4 in Rhabdomyosarcoma with the Inhibitor Ponatinib (AP24534)

    doi: 10.1371/journal.pone.0076551

    Figure Lengend Snippet: In vivo tumor growth assay with daily treatment of 30 mg/kg of ponatinib after tumor volumes reach 100 mm 3 . Arrow indicates the start of treatment. (A) Treatment of tumors harboring the FGFR4 N535K mutation with ponatinib significantly inhibits tumor growth after 10 days of treatment (*p = 0.0165, **p = 0.0048). (B) Treatment of tumors containing the FGFR4 V550E mutation with ponatinib significantly inhibits tumor growth after 6 days of treatment (*p = 0.0185, **p = 0.0087, ***p = 0.0005). (C) Treatment of tumors expressing the wild-type FGFR4 with ponatinib does not affect tumor growth. (D) Treatment of tumors expressing the empty vector with ponatinib does not affect tumor growth.

    Article Snippet: Quantitative RT-PCR using Taqman assays (FGFR4: Hs00242558_m1 and GAPDH: Hs99999905_m1) on a Fluidigm system was previously described .

    Techniques: In Vivo, Growth Assay, Mutagenesis, Expressing, Plasmid Preparation

    Western blot analysis of expression and phosphorylation of wild-type and mutated FGFR4 and its downstream target, STAT3, after treatment with 0, 200, and 800 nM concentrations of ponatinib (AP24534) for 8 hours. (A) A dose-dependent decrease in wild-type FGFR4 phosphorylation as shown by immunoprecipitation of FGFR4 and immunoblotting for phosphotyrosine. (B) A similar dose-dependent inhibition is seen for FGFR4 with the V550E and N535K mutation. (C-D) Western blot shows a dose-dependent decrease in STAT3 phosphorylation after treatment with ponatinib for three fusion-positive (RH4, RH5, and RH41) and one fusion-negative (CTR) RMS cell lines as well as the two RMS772 cell lines expressing the FGFR4 mutations N535K and V550E.

    Journal: PLoS ONE

    Article Title: Targeting Wild-Type and Mutationally Activated FGFR4 in Rhabdomyosarcoma with the Inhibitor Ponatinib (AP24534)

    doi: 10.1371/journal.pone.0076551

    Figure Lengend Snippet: Western blot analysis of expression and phosphorylation of wild-type and mutated FGFR4 and its downstream target, STAT3, after treatment with 0, 200, and 800 nM concentrations of ponatinib (AP24534) for 8 hours. (A) A dose-dependent decrease in wild-type FGFR4 phosphorylation as shown by immunoprecipitation of FGFR4 and immunoblotting for phosphotyrosine. (B) A similar dose-dependent inhibition is seen for FGFR4 with the V550E and N535K mutation. (C-D) Western blot shows a dose-dependent decrease in STAT3 phosphorylation after treatment with ponatinib for three fusion-positive (RH4, RH5, and RH41) and one fusion-negative (CTR) RMS cell lines as well as the two RMS772 cell lines expressing the FGFR4 mutations N535K and V550E.

    Article Snippet: Quantitative RT-PCR using Taqman assays (FGFR4: Hs00242558_m1 and GAPDH: Hs99999905_m1) on a Fluidigm system was previously described .

    Techniques: Western Blot, Expressing, Immunoprecipitation, Inhibition, Mutagenesis

    RMS cell lines with overexpressed FGFR4 are more sensitive to ponatinib. (A) The sensitivity of a panel of six fusion-positive RMS cell lines (RH4, RH28, JR, RH41, RH5, and RH30) and eight fusion-negative RMS cell lines (BIRCH, RH18, TTC-442, CT-10, CTR, TTC-516, RD, and RH36) to ponatinib is correlated to FGFR4 mRNA expression levels by Spearman ranking (p = 0.0261). (B) Comparing the variation in IC 50 values of fusion-positive (FP) and fusion-negative (FN) RMS cell lines shows a significant difference by F test (p = 0.0125). (C) A difference in IC 50 values can be seen between RMS cell lines expressing low (below a relative level of 6) and high (above a relative level of 6) levels of FGFR4 (p = 0.0344).

    Journal: PLoS ONE

    Article Title: Targeting Wild-Type and Mutationally Activated FGFR4 in Rhabdomyosarcoma with the Inhibitor Ponatinib (AP24534)

    doi: 10.1371/journal.pone.0076551

    Figure Lengend Snippet: RMS cell lines with overexpressed FGFR4 are more sensitive to ponatinib. (A) The sensitivity of a panel of six fusion-positive RMS cell lines (RH4, RH28, JR, RH41, RH5, and RH30) and eight fusion-negative RMS cell lines (BIRCH, RH18, TTC-442, CT-10, CTR, TTC-516, RD, and RH36) to ponatinib is correlated to FGFR4 mRNA expression levels by Spearman ranking (p = 0.0261). (B) Comparing the variation in IC 50 values of fusion-positive (FP) and fusion-negative (FN) RMS cell lines shows a significant difference by F test (p = 0.0125). (C) A difference in IC 50 values can be seen between RMS cell lines expressing low (below a relative level of 6) and high (above a relative level of 6) levels of FGFR4 (p = 0.0344).

    Article Snippet: Quantitative RT-PCR using Taqman assays (FGFR4: Hs00242558_m1 and GAPDH: Hs99999905_m1) on a Fluidigm system was previously described .

    Techniques: Expressing

    RMS772 cell harboring activating FGFR4 mutations V550E or N535K are more sensitive to ponatinib (AP24534) after 24 hour treatment than RMS772 cells expressing wild-type (WT) FGFR4 or the empty vector (VCtrl) (*p =

    Journal: PLoS ONE

    Article Title: Targeting Wild-Type and Mutationally Activated FGFR4 in Rhabdomyosarcoma with the Inhibitor Ponatinib (AP24534)

    doi: 10.1371/journal.pone.0076551

    Figure Lengend Snippet: RMS772 cell harboring activating FGFR4 mutations V550E or N535K are more sensitive to ponatinib (AP24534) after 24 hour treatment than RMS772 cells expressing wild-type (WT) FGFR4 or the empty vector (VCtrl) (*p =

    Article Snippet: Quantitative RT-PCR using Taqman assays (FGFR4: Hs00242558_m1 and GAPDH: Hs99999905_m1) on a Fluidigm system was previously described .

    Techniques: Expressing, Plasmid Preparation

    Ponatinib (AP24534) holds cell cycling at sub G 1 phase and induces cell death via apoptosis. (A) Cell cycle analysis of the two most sensitive cell lines to ponatinib, RH4 and RH5, and the two RMS772 cell lines expressing FGFR4 mutations (N535K and V550E) showed increased time in sub G 1 phase and decreased time in S phase across all four cell lines after 24 hours of treatment with 2.5 µM ponatinib. (B) Cell death induced by 2.5 µM ponatinib treatment is mediated via the caspase 3/7 pathway (*p = 0.0029, **p = 0.0027, ***p = 0.0017, ****p = 0.0001).

    Journal: PLoS ONE

    Article Title: Targeting Wild-Type and Mutationally Activated FGFR4 in Rhabdomyosarcoma with the Inhibitor Ponatinib (AP24534)

    doi: 10.1371/journal.pone.0076551

    Figure Lengend Snippet: Ponatinib (AP24534) holds cell cycling at sub G 1 phase and induces cell death via apoptosis. (A) Cell cycle analysis of the two most sensitive cell lines to ponatinib, RH4 and RH5, and the two RMS772 cell lines expressing FGFR4 mutations (N535K and V550E) showed increased time in sub G 1 phase and decreased time in S phase across all four cell lines after 24 hours of treatment with 2.5 µM ponatinib. (B) Cell death induced by 2.5 µM ponatinib treatment is mediated via the caspase 3/7 pathway (*p = 0.0029, **p = 0.0027, ***p = 0.0017, ****p = 0.0001).

    Article Snippet: Quantitative RT-PCR using Taqman assays (FGFR4: Hs00242558_m1 and GAPDH: Hs99999905_m1) on a Fluidigm system was previously described .

    Techniques: Cell Cycle Assay, Expressing

    N-cadherin is involved in the pro-oncogenic role of Arg388 FGFR4 overexpression. ( A ) N-cadherin silencing in the FGFR4-388Arg-overexpressing H2009 cell line using a shRNA approach. Clonability ( B ) and soft agar assays ( C ) of the FGFR4-388Arg-overexpressing, N-cadherin silenced H2009 cell line. ( D ) Relative tumor growth of the xenograft in immunodeprived nude mice of these cell lines. ( E ) Western blot showing the STAT3 activation suspension by the STAT3 inhibitor SI3-201 along the time, which is accompanied by N-cadherin protein levels reduction, in the FGFR4-388Arg-overexpressing H2009 and H226 cell lines. SI3-201 exposure in hours is indicated in the figure. ( F ) Western blot showing the effect of STAT3 inhibition and N-cadherin shRNA silencing in several cancer-related downstream signaling pathways in the EV, FGFR4-388Gly and -388Arg-overexpressing H2009 cell line. In soft agar and clonability assays, colony number representation is shown. All values were normalized to empty vector control for each replicate of the experiment and the mean and standard deviation of every normalized replicate are represented. For western blots, a representative blot is shown. EV = Empty Vector control, FGFR4-Gly = FGFR4-388Gly overexpressing, FGFR4-Arg = FGFR4-388Arg overexpressing. p-Values are represented as asterisks (*p

    Journal: Scientific Reports

    Article Title: The FGFR4-388arg Variant Promotes Lung Cancer Progression by N-Cadherin Induction

    doi: 10.1038/s41598-018-20570-3

    Figure Lengend Snippet: N-cadherin is involved in the pro-oncogenic role of Arg388 FGFR4 overexpression. ( A ) N-cadherin silencing in the FGFR4-388Arg-overexpressing H2009 cell line using a shRNA approach. Clonability ( B ) and soft agar assays ( C ) of the FGFR4-388Arg-overexpressing, N-cadherin silenced H2009 cell line. ( D ) Relative tumor growth of the xenograft in immunodeprived nude mice of these cell lines. ( E ) Western blot showing the STAT3 activation suspension by the STAT3 inhibitor SI3-201 along the time, which is accompanied by N-cadherin protein levels reduction, in the FGFR4-388Arg-overexpressing H2009 and H226 cell lines. SI3-201 exposure in hours is indicated in the figure. ( F ) Western blot showing the effect of STAT3 inhibition and N-cadherin shRNA silencing in several cancer-related downstream signaling pathways in the EV, FGFR4-388Gly and -388Arg-overexpressing H2009 cell line. In soft agar and clonability assays, colony number representation is shown. All values were normalized to empty vector control for each replicate of the experiment and the mean and standard deviation of every normalized replicate are represented. For western blots, a representative blot is shown. EV = Empty Vector control, FGFR4-Gly = FGFR4-388Gly overexpressing, FGFR4-Arg = FGFR4-388Arg overexpressing. p-Values are represented as asterisks (*p

    Article Snippet: Preamplification and gene expression analysis were conducted using TaqMan probes from Life Technologies: Hs01106908_m1 FAM (FGFR4), Hs00983056_m1 FAM (N-cadherin), and Hs99999905_m1 FAM (GAPDH).

    Techniques: Over Expression, shRNA, Mouse Assay, Western Blot, Activation Assay, Inhibition, Plasmid Preparation, Standard Deviation

    Overexpression of the 388Arg variant of FGFR4 induces the expression of EMT markers in lung cell lines. The mRNA ( A ) and protein ( B ) expression of four EMT markers (N-cadherin, vimentin, Twist1 and Snail1) was performed in the control empty vector (EV), FGFR4-388Gly-overexpressing (FGFR4-Gly) and FGFR4-388Arg-overexpressing (FGFR4-Arg) lung immortalized NL20 cell line and lung SCC H226 and Calu1, and ADC H2009 and HCC827 cell lines. ( C ) Relative number of migrated cells of the previously mentioned cell lines. The mRNA measurements were performed in three independent experiments and mean expression values, represented as 2 −ΔCt with their respective standard deviation, are represented. For western blots, a representative blot is shown. Migration assays were performed in three independent experiments. All values were normalized to empty vector control for each replicate of the experiment and the mean and standard deviation of every normalized replicate are represented. p-values are represented as asterisks (*p

    Journal: Scientific Reports

    Article Title: The FGFR4-388arg Variant Promotes Lung Cancer Progression by N-Cadherin Induction

    doi: 10.1038/s41598-018-20570-3

    Figure Lengend Snippet: Overexpression of the 388Arg variant of FGFR4 induces the expression of EMT markers in lung cell lines. The mRNA ( A ) and protein ( B ) expression of four EMT markers (N-cadherin, vimentin, Twist1 and Snail1) was performed in the control empty vector (EV), FGFR4-388Gly-overexpressing (FGFR4-Gly) and FGFR4-388Arg-overexpressing (FGFR4-Arg) lung immortalized NL20 cell line and lung SCC H226 and Calu1, and ADC H2009 and HCC827 cell lines. ( C ) Relative number of migrated cells of the previously mentioned cell lines. The mRNA measurements were performed in three independent experiments and mean expression values, represented as 2 −ΔCt with their respective standard deviation, are represented. For western blots, a representative blot is shown. Migration assays were performed in three independent experiments. All values were normalized to empty vector control for each replicate of the experiment and the mean and standard deviation of every normalized replicate are represented. p-values are represented as asterisks (*p

    Article Snippet: Preamplification and gene expression analysis were conducted using TaqMan probes from Life Technologies: Hs01106908_m1 FAM (FGFR4), Hs00983056_m1 FAM (N-cadherin), and Hs99999905_m1 FAM (GAPDH).

    Techniques: Over Expression, Variant Assay, Expressing, Plasmid Preparation, Standard Deviation, Western Blot, Migration

    Arg388 FGFR4 mRNA expression correlates with N-cadherin mRNA expression. ( A ) N-cadherin mRNA expression levels according to Arg388 FGFR4 mRNA expression. ( B ) Bivariate correlation analysis of Arg388 FGFR4 and N-cadherin mRNA expression levels in the whole NSCLC cohort and in the ADC and SCC patient subsets. Gly = FGFR4-388Gly, Arg = FGFR4-388Arg.

    Journal: Scientific Reports

    Article Title: The FGFR4-388arg Variant Promotes Lung Cancer Progression by N-Cadherin Induction

    doi: 10.1038/s41598-018-20570-3

    Figure Lengend Snippet: Arg388 FGFR4 mRNA expression correlates with N-cadherin mRNA expression. ( A ) N-cadherin mRNA expression levels according to Arg388 FGFR4 mRNA expression. ( B ) Bivariate correlation analysis of Arg388 FGFR4 and N-cadherin mRNA expression levels in the whole NSCLC cohort and in the ADC and SCC patient subsets. Gly = FGFR4-388Gly, Arg = FGFR4-388Arg.

    Article Snippet: Preamplification and gene expression analysis were conducted using TaqMan probes from Life Technologies: Hs01106908_m1 FAM (FGFR4), Hs00983056_m1 FAM (N-cadherin), and Hs99999905_m1 FAM (GAPDH).

    Techniques: Expressing

    Effect of the overexpression of the 388Gly and 388Arg variants of FGFR4 in the tumorigenic abilities of the inmortalized NL20, the lung SCC H226 and Calu-1, and the H2009 and HCC827 lung ADC cell lines. ( A ) 10% FBS growth curves. ( B ) soft agar assays showing the relative colony number quantification (left) and representative images (right). ( C ) Western blot determination of activation of cancer-related signalling pathways of 388Gly and 388Arg FGFR4-overexpressing cell lines. In soft agar assay, colony number representation is shown. All values were normalized to empty vector control for each replicate of the experiment, and the mean and standard deviation of every normalized replicate are represented. For western blotting, cells were serum starved for 5 hours and then the protein extraction was made. For the serum stimulated conditions, after serum starvation cells were stimulated with serum-containing complete medium for fifteen minutes before protein extraction. p-Values are represented as asterisks (*p

    Journal: Scientific Reports

    Article Title: The FGFR4-388arg Variant Promotes Lung Cancer Progression by N-Cadherin Induction

    doi: 10.1038/s41598-018-20570-3

    Figure Lengend Snippet: Effect of the overexpression of the 388Gly and 388Arg variants of FGFR4 in the tumorigenic abilities of the inmortalized NL20, the lung SCC H226 and Calu-1, and the H2009 and HCC827 lung ADC cell lines. ( A ) 10% FBS growth curves. ( B ) soft agar assays showing the relative colony number quantification (left) and representative images (right). ( C ) Western blot determination of activation of cancer-related signalling pathways of 388Gly and 388Arg FGFR4-overexpressing cell lines. In soft agar assay, colony number representation is shown. All values were normalized to empty vector control for each replicate of the experiment, and the mean and standard deviation of every normalized replicate are represented. For western blotting, cells were serum starved for 5 hours and then the protein extraction was made. For the serum stimulated conditions, after serum starvation cells were stimulated with serum-containing complete medium for fifteen minutes before protein extraction. p-Values are represented as asterisks (*p

    Article Snippet: Preamplification and gene expression analysis were conducted using TaqMan probes from Life Technologies: Hs01106908_m1 FAM (FGFR4), Hs00983056_m1 FAM (N-cadherin), and Hs99999905_m1 FAM (GAPDH).

    Techniques: Over Expression, Western Blot, Activation Assay, Soft Agar Assay, Plasmid Preparation, Standard Deviation, Protein Extraction

    Correlation of FGFR4 variant and prognosis in high FGFR4 mRNA expressing NSCLC patients. ( A ) Overall and progression-free survival curves for high FGFR4 mRNA expressing patients, according to the FGFR4 variant in the whole NSCLC cohort. ( B ) Overall and progression-free survival analysis of SCC and ADC patient subsets depending on the FGFR4 variant, taking into account exclusively the groups with high FGFR4 mRNA expression. Gly = FGFR4-388Gly, Arg = FGFR4-388Arg.

    Journal: Scientific Reports

    Article Title: The FGFR4-388arg Variant Promotes Lung Cancer Progression by N-Cadherin Induction

    doi: 10.1038/s41598-018-20570-3

    Figure Lengend Snippet: Correlation of FGFR4 variant and prognosis in high FGFR4 mRNA expressing NSCLC patients. ( A ) Overall and progression-free survival curves for high FGFR4 mRNA expressing patients, according to the FGFR4 variant in the whole NSCLC cohort. ( B ) Overall and progression-free survival analysis of SCC and ADC patient subsets depending on the FGFR4 variant, taking into account exclusively the groups with high FGFR4 mRNA expression. Gly = FGFR4-388Gly, Arg = FGFR4-388Arg.

    Article Snippet: Preamplification and gene expression analysis were conducted using TaqMan probes from Life Technologies: Hs01106908_m1 FAM (FGFR4), Hs00983056_m1 FAM (N-cadherin), and Hs99999905_m1 FAM (GAPDH).

    Techniques: Variant Assay, Expressing

    FGFR4 80-bp deletion line lost FGFR4 expression and signaling. A , genotyping characterization of the 80-bp deletion FGFR4 KO animals. A predicted 279-bp band in WT littermate control animals, 199 bp in KO animals, and both bands in heterozygous ( HET )

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: FGFR4 80-bp deletion line lost FGFR4 expression and signaling. A , genotyping characterization of the 80-bp deletion FGFR4 KO animals. A predicted 279-bp band in WT littermate control animals, 199 bp in KO animals, and both bands in heterozygous ( HET )

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques: Expressing

    FGFR4 KO mice maintained response to FGF19 treatment induced changes in glucose and insulin regulation. 9–11-week-old male WT and KO mice were fed with 60 kcal % high fat diet for 8 weeks. A , OGTT responses of WT and FGFR4 KO animals ( n = 25 each

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: FGFR4 KO mice maintained response to FGF19 treatment induced changes in glucose and insulin regulation. 9–11-week-old male WT and KO mice were fed with 60 kcal % high fat diet for 8 weeks. A , OGTT responses of WT and FGFR4 KO animals ( n = 25 each

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques: Mouse Assay

    Improved glucose metabolism and insulin sensitivity in FGFR4 KO animals upon high fat diet feeding. 12-Week-old male WT and KO mice were fed with 60 kcal % high fat diet. Body weight ( A ), OGTT glucose response ( B ), serum insulin ( C ), and fasting glucose

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: Improved glucose metabolism and insulin sensitivity in FGFR4 KO animals upon high fat diet feeding. 12-Week-old male WT and KO mice were fed with 60 kcal % high fat diet. Body weight ( A ), OGTT glucose response ( B ), serum insulin ( C ), and fasting glucose

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques: Mouse Assay

    FGFR4 KO Mice Display Significant Improvement in Glucose Metabolism and Insulin Sensitivity

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: FGFR4 KO Mice Display Significant Improvement in Glucose Metabolism and Insulin Sensitivity

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques: Mouse Assay

    Metabolic parameters were not significantly changed in FGFR4 KO animals upon chow diet feeding. Body weight ( A ), serum insulin ( B ), serum triglyceride ( C ), and OGTT glucose response ( D ) were recorded from 13–14-week-old age-matched WT and KO animals

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: Metabolic parameters were not significantly changed in FGFR4 KO animals upon chow diet feeding. Body weight ( A ), serum insulin ( B ), serum triglyceride ( C ), and OGTT glucose response ( D ) were recorded from 13–14-week-old age-matched WT and KO animals

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques:

    FGFR4 KO Mice Display Significant Improvement in Glucose Metabolism and Insulin Sensitivity

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: FGFR4 KO Mice Display Significant Improvement in Glucose Metabolism and Insulin Sensitivity

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques: Mouse Assay

    A model of FGFR4-mediated metabolic regulations. FGFR4 regulates bile acid synthesis by suppressing expression of Cyp7a1 , the bile acid-producing enzyme in liver. In FGFR4 KO, increased basal bile acid synthesis triggered up-regulation of FGF21 in liver

    Journal:

    Article Title: Fibroblast Growth Factor Receptor 4 (FGFR4) Deficiency Improves Insulin Resistance and Glucose Metabolism under Diet-induced Obesity Conditions

    doi: 10.1074/jbc.M114.592022

    Figure Lengend Snippet: A model of FGFR4-mediated metabolic regulations. FGFR4 regulates bile acid synthesis by suppressing expression of Cyp7a1 , the bile acid-producing enzyme in liver. In FGFR4 KO, increased basal bile acid synthesis triggered up-regulation of FGF21 in liver

    Article Snippet: FGFR4 primers from ABI (Applied Biosystems, Mm01341852_m1 located in exon boundary 11–12) were also used. qRT-PCR was performed on the Stratagene Mx3000P quantitative PCR machine with the Stratagene Brilliant II qRT-PCR Master Mix Kit, 1-Step (600809, Stratagene) using 50 ng of RNA/well and normalized to mouse GAPDH (4352932E and 4352339E, ABI).

    Techniques: Expressing