Journal: BMJ Open

Article Title: Association between serum vitamin B 6 concentration and risk of osteoporosis in the middle-aged and older people in China: a cross-sectional study

doi: 10.1136/bmjopen-2018-028129

Figure Lengend Snippet: Comparison of serum Vit B 6 and serological metabolism parameters among control, osteopenia and osteoporosis groups

Article Snippet: The concentration of fibroblast growth factor 23 (FGF23) was measured through an ELISA performed using a human FGF23 ELISA kit (CSB-E10113h, Cusabio Biotech, Wuhan, China) and a micro plate reader (MK3, Thermo, Waltham, Massachusetts, USA).

Techniques: Comparison, Control

Journal: BMJ Open

Article Title: Association between serum vitamin B 6 concentration and risk of osteoporosis in the middle-aged and older people in China: a cross-sectional study

doi: 10.1136/bmjopen-2018-028129

Figure Lengend Snippet: Relationship between serum Vit B 6 concentrations and bone metabolism parameter concentrations in control, osteopenia and osteoporosis groups

Article Snippet: The concentration of fibroblast growth factor 23 (FGF23) was measured through an ELISA performed using a human FGF23 ELISA kit (CSB-E10113h, Cusabio Biotech, Wuhan, China) and a micro plate reader (MK3, Thermo, Waltham, Massachusetts, USA).

Techniques: Control

Journal: Medicina (Kaunas, Lithuania)

Article Title: Fibroblast Growth Factor 23 Signaling Does Not Increase Inflammation from Pseudomonas aeruginosa Infection in the Cystic Fibrosis Bronchial Epithelium.

doi: 10.3390/medicina59091635

Figure Lengend Snippet: Figure 1. FGF23, in combination with PA infection, did not augment transcript expression of pro- inflammatory cytokines when compared to PA infection alone. (A) Fold change in mRNA levels of FGFRs 1–4 with or without FGF23 and PA (PAM57-15). (B) Fold change in mRNA levels of CXCL8 (IL-8), IL-6, and IL-1β after treatment with or without FGF23 and with or without PA. (C) Fold change in mRNA levels of TGFβ and klotho after treatment with or without FGF23 with or without PA infection. Data was represented as fold change in mRNA expression with n = 3 independent experiments. Statistical analysis was performed using a 2-way ANOVA, followed by Tukey’s multiple comparison post hoc test. Data are expressed as means ± standard error of the mean (SEM). Abbrv. ns = not significant. Differences were considered statistically significant if * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: Where indicated, human recombinant FGF23 (40 ng/mL, RnD Systems, Minneapolis, MN, USA, 2604FG), AZD4547 (10 μM, Selleckchem, Houston, TX, USA, S2801) for (FGFR) 1–3 inhibition, or BLU9931 (0.1 μM, Selleckchem, S7819) for FGFR4 inhibition were used as a pre-treatment for 1 h before infection with or without PA.

Techniques: Infection, Expressing, Comparison, IF-P

Journal: Medicina (Kaunas, Lithuania)

Article Title: Fibroblast Growth Factor 23 Signaling Does Not Increase Inflammation from Pseudomonas aeruginosa Infection in the Cystic Fibrosis Bronchial Epithelium.

doi: 10.3390/medicina59091635

Figure Lengend Snippet: Figure 2. FGF23 did not alter pro-inflammatory marker production and secretion in PA-infected CFBEs. (A) Graph showing levels of IL-8 protein expression in the basolateral media, cell lysate, and apical media when CFBEs were treated with or without FGF23 and with or without PA infection. (B) Graphs showing levels of IL-6 protein expression in the basolateral media, cell lysate, and apical media when CFBEs were treated with or without FGF23 and with or without PA infection. Data was represented as expression (pg/mL) with n = 5 independent experiments. Statistical analysis was performed using a 2-way ANOVA, followed by Tukey’s multiple comparison post hoc test. Data are expressed as means ± standard error of the mean (SEM). Abbrv. ns = not significant. Differences were considered statistically significant if * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: Where indicated, human recombinant FGF23 (40 ng/mL, RnD Systems, Minneapolis, MN, USA, 2604FG), AZD4547 (10 μM, Selleckchem, Houston, TX, USA, S2801) for (FGFR) 1–3 inhibition, or BLU9931 (0.1 μM, Selleckchem, S7819) for FGFR4 inhibition were used as a pre-treatment for 1 h before infection with or without PA.

Techniques: Marker, Infection, Expressing, Comparison, IF-P

Journal: Medicina (Kaunas, Lithuania)

Article Title: Fibroblast Growth Factor 23 Signaling Does Not Increase Inflammation from Pseudomonas aeruginosa Infection in the Cystic Fibrosis Bronchial Epithelium.

doi: 10.3390/medicina59091635

Figure Lengend Snippet: Figure 3. FGF23 did not alter the phosphorylation of PLCγ or ERK in the context of PA infection in CFBEs. (A) Quantification of Western Blots for phospho−ERK and phospho−PLCγ of PA-infected CFBEs compared to uninfected control CFBEs. (B) Representative western blots for phospho−PLCγ, total PLCγ, phospho−ERK, and total ERK with or without FGF23 treatment and with or without PA infection with the mucoid strain PAM57-15. (C) Densitometric quantification of Western Blot images from panel (B). Data was represented as phosphorylated PLCγ or ERK over their respective total, also normalized to total protein via the quantification of the Coomassie gel. These graphs represent n = 5−6 independent experiments. Statistical analysis was performed using a 1−way or 2−way ANOVA, followed by the Kruskal−Wallis test or Tukey’s multiple comparison post hoc test, respectively. Abbrv. ns = not significant. Data are expressed as means ± standard error of the mean (SEM). Differences were considered statistically significant if * p < 0.05.

Article Snippet: Where indicated, human recombinant FGF23 (40 ng/mL, RnD Systems, Minneapolis, MN, USA, 2604FG), AZD4547 (10 μM, Selleckchem, Houston, TX, USA, S2801) for (FGFR) 1–3 inhibition, or BLU9931 (0.1 μM, Selleckchem, S7819) for FGFR4 inhibition were used as a pre-treatment for 1 h before infection with or without PA.

Techniques: Phospho-proteomics, Infection, Western Blot, Control, Comparison, IF-P

Journal: International Journal of Molecular Sciences

Article Title: FGF23-Mediated Activation of Local RAAS Promotes Cardiac Hypertrophy and Fibrosis

doi: 10.3390/ijms20184634

Figure Lengend Snippet: Characteristics of sham and 5/6 nephrectomized (5/6Nx) rats.

Article Snippet: The next day, NRVM and NRCF were stimulated for 48 h with recombinant human FGF23 (NRVM, 100 ng/mL; NRCF, 10 ng/mL; R&D Systems, Wiesbaden, Germany) alone and in combination with 100 nM cyclosporine A (CsA), 1 μM losartan (Los) or 10 μM spironolactone (Spiro).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: FGF23-Mediated Activation of Local RAAS Promotes Cardiac Hypertrophy and Fibrosis

doi: 10.3390/ijms20184634

Figure Lengend Snippet: The 5/6 nephrectomized (5/6Nx) rats develop left ventricular (LV) fibrosis, which correlates with fibroblast growth factor 23 ( Fgf23 ) expression. ( A ) Representative bright field and polarized light microscopy images of picrosirius red-stained myocardial tissue of 5/6Nx and sham-operated rats and quantification of interstitial collagen fiber deposition demonstrating increased LV fibrosis in 5/6Nx rats (scale bar, 50 µm). ( B–D ) Pearson’s correlations of LV fibrosis with cross-sectional area of cardiomyocytes, and cardiac and bone Fgf23 mRNA expression as determined by quantitative real-time PCR using Gapdh as housekeeping gene. Clear dots, sham-operated rats; black dots, 5/6Nx rats. All values are shown as mean ± SEM; ** p < 0.01; n = 5–6 rats per group.

Article Snippet: The next day, NRVM and NRCF were stimulated for 48 h with recombinant human FGF23 (NRVM, 100 ng/mL; NRCF, 10 ng/mL; R&D Systems, Wiesbaden, Germany) alone and in combination with 100 nM cyclosporine A (CsA), 1 μM losartan (Los) or 10 μM spironolactone (Spiro).

Techniques: Expressing, Light Microscopy, Staining, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: FGF23-Mediated Activation of Local RAAS Promotes Cardiac Hypertrophy and Fibrosis

doi: 10.3390/ijms20184634

Figure Lengend Snippet: Here, 5/6 nephrectomy (5/6Nx) in rats induces cardiac expression of renin-angiotensin-aldosterone system (RAAS)-associated genes that correlates with left ventricular (LV) fibrosis. ( A–C ) Quantitative real-time PCR analyses in 5/6Nx rats show cardiac-specific induction of angiotensinogen ( Agt ), renin ( Ren ) and angiotensin converting enzyme ( Ace ), while ( D ) cardiac angiotensin II receptor type 1 ( AT1R ) mRNA expression is not significantly induced. ( E–G ) Pearson’s correlations of LV fibrosis with cardiac expression of Agt , Ace and AT1R , and ( H ) of cardiac Fgf23 with Agt mRNA expression. Clear dots, sham-operated rats; black dots, 5/6Nx rats. All values are shown as mean ± SEM; * p < 0.05, *** p < 0.001; n = 5–6 rats per group.

Article Snippet: The next day, NRVM and NRCF were stimulated for 48 h with recombinant human FGF23 (NRVM, 100 ng/mL; NRCF, 10 ng/mL; R&D Systems, Wiesbaden, Germany) alone and in combination with 100 nM cyclosporine A (CsA), 1 μM losartan (Los) or 10 μM spironolactone (Spiro).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: FGF23-Mediated Activation of Local RAAS Promotes Cardiac Hypertrophy and Fibrosis

doi: 10.3390/ijms20184634

Figure Lengend Snippet: FGF23 induces RAAS-associated genes in neonatal rat ventricular myocytes (NRVM) and cardiac fibroblasts (NRCF) in vitro as shown by quantitative real-time PCR. ( A–D ) In NRVM, FGF23 treatment increases the mRNA expression of angiotensinogen ( Agt ) and renin ( Ren ), whereas the effects of FGF23 on angiotensin converting enzyme ( Ace ) and angiotensin II receptor type 1 ( AT1R ) expression levels are not statistically significant. ( E ) FGF23 and aldosterone (Aldo) induce the mRNA expression of neutrophil gelatinase-associated lipocalin ( Ngal ) in NRVM. ( F–I ) In NRCF, Agt , Ace and Ren mRNA expressions are enhanced after FGF23 treatment, but FGF23 does not significantly induce AT1R . ( J ) Ngal mRNA expression is elevated upon FGF23 and aldosterone stimulation. All values are shown as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control; n = 6–8 independent cell isolations.

Article Snippet: The next day, NRVM and NRCF were stimulated for 48 h with recombinant human FGF23 (NRVM, 100 ng/mL; NRCF, 10 ng/mL; R&D Systems, Wiesbaden, Germany) alone and in combination with 100 nM cyclosporine A (CsA), 1 μM losartan (Los) or 10 μM spironolactone (Spiro).

Techniques: In Vitro, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: FGF23-Mediated Activation of Local RAAS Promotes Cardiac Hypertrophy and Fibrosis

doi: 10.3390/ijms20184634

Figure Lengend Snippet: FGF23-induced hypertrophy in cultured NRVM is prevented by cyclosporine A (CsA), losartan (Los) and spironolactone (Spiro). ( A ) Representative immunofluorescent images of NRVM stained with sarcomeric α-actinin (green) and DAPI (blue) (scale bar, 50 µm), and quantification of NRVM size show significant induction of hypertrophic growth after treatment with FGF23, which is ameliorated by co-treatment with CsA, Los and Spiro. Phenylephrine (PE) served as positive control. ( B , C ) Quantitative real-time PCR analyses reveal FGF23-mediated increase of mRNA expression of prohypertrophic markers ANP and BNP, which is inhibited by CsA, Los and Spiro. Gapdh served as housekeeping gene. All values are shown as mean ± SEM; ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control; § p < 0.05, §§§§ p < 0.0001 versus FGF23; n = 5–9 independent cell isolations.

Article Snippet: The next day, NRVM and NRCF were stimulated for 48 h with recombinant human FGF23 (NRVM, 100 ng/mL; NRCF, 10 ng/mL; R&D Systems, Wiesbaden, Germany) alone and in combination with 100 nM cyclosporine A (CsA), 1 μM losartan (Los) or 10 μM spironolactone (Spiro).

Techniques: Cell Culture, Staining, Positive Control, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: FGF23-Mediated Activation of Local RAAS Promotes Cardiac Hypertrophy and Fibrosis

doi: 10.3390/ijms20184634

Figure Lengend Snippet: FGF23 stimulates the proliferation of NRCF and induces the expression of the profibrotic markers collagen 1 ( Col1 ), transforming growth factor beta 1 ( Tgfb ) and connective tissue growth factor ( Ctgf ). ( A ) The proliferation of NRCF increases in response to stimulation with FGF23, which was not inhibited in the presence of cyclosporine A (CsA), losartan (Los) or spironolactone (Spiro) as demonstrated by MTS-based proliferation assay. ( B–D ) The mRNA expression of Tgfb, Ctgf and Col1 in NRCF is increased upon FGF23 stimulation as measured by quantitative real-time PCR. Co-treatment with Los and Spiro inhibits the induction of fibrotic markers by FGF23, while CsA has lesser effects. TGFβ and angiotensin II (AngII) served as positive controls. Horizontal dotted lines represent the level of controls. All values are shown as mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control; § p < 0.05 versus FGF23; n = 5–9 independent cell isolations.

Article Snippet: The next day, NRVM and NRCF were stimulated for 48 h with recombinant human FGF23 (NRVM, 100 ng/mL; NRCF, 10 ng/mL; R&D Systems, Wiesbaden, Germany) alone and in combination with 100 nM cyclosporine A (CsA), 1 μM losartan (Los) or 10 μM spironolactone (Spiro).

Techniques: Expressing, Proliferation Assay, Real-time Polymerase Chain Reaction, Control

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Dependence of the SPRi signal on FGF23 antibody concentration. The used concentration of the FGF23-αKlotho complex was 40:169.2 pg/mL; pH = 7.4. “Standard deviation (SD)” was calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Dependence of the SPRi signal on the FGF23 concentration. The used FGF23 antibody concentration is 25 ng/mL; pH = 7.4. SD was calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Dependence of the SPRi signal on the FGF23 concentration with the addition of αKlotho protein. The used FGF23 antibody concentration is 25 ng/mL; pH = 7.4. SD was calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Influence of the αKlotho protein on the detected FGF23 concentration. SD was calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Influence of the time on the formation of the FGF23-αKlotho complex. SD was calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: The influence of the potential interferents on the determination of the FGF23 with array SPRi biosensor. SD was calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Precision and accuracy of the FGF23 SPRi biosensor. SD and RSD were calculated from 36 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Precision and recovery of the FGF23 SPRi biosensor tested in a plasma sample after the addition of a 25 pg/mL spike. SD and RSD were calculated from 12 individual measurements.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Clinical Proteomics

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Formation of the FGF23 SPRi biosensor layers, i.e., bare gold, cysteamine, FGF23 antibody, and FGF23-αKlotho complex, captured via SEM. Used parameters: 12.5 or 15 kV, and 100,000× magnification.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: The comparison of the results of FGF23 concentration measurements performed with the SPRi biosensor and ELISA in blood plasma samples. SD was calculated based on 12 measurements for the SPRi biosensor and 2 for the ELISA.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques: Comparison, Concentration Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: International Journal of Molecular Sciences

Article Title: A New Approach to the Quantification of Fibroblast Growth Factor 23—An Array Surface Plasmon Resonance Imaging Biosensor

doi: 10.3390/ijms242015327

Figure Lengend Snippet: Scheme of the measuring system of the SPRi apparatus and an overview diagram of the FGF23 biosensor.

Article Snippet: The monoclonal mouse anti-human FGF23 antibody (R&D Systems) was a receptor that bound the FGF23 protein from samples to the chip’s surface.

Techniques:

Journal: Frontiers in Nephrology

Article Title: Role of kidney transplantation in long-term cardiac reverse remodeling and interconnecting mechanisms in type 4 cardiorenal syndrome

doi: 10.3389/fneph.2024.1455036

Figure Lengend Snippet: (A) displays the distribution of post-KT FGF23 levels based on the hypertension status. (B) shows that the patients with larger LA volumes had the worst NHYA class.

Article Snippet: FGF23 levels were quantified using the Human FGF23 monoclonal antibody and R&D Systems reagent (Minneapolis, MN, USA).

Techniques:

Journal: Frontiers in Nephrology

Article Title: Role of kidney transplantation in long-term cardiac reverse remodeling and interconnecting mechanisms in type 4 cardiorenal syndrome

doi: 10.3389/fneph.2024.1455036

Figure Lengend Snippet: Biochemical and echocardiographic characteristics grouped according the basal LVEF and after KT.

Article Snippet: FGF23 levels were quantified using the Human FGF23 monoclonal antibody and R&D Systems reagent (Minneapolis, MN, USA).

Techniques:

Journal: PLoS ONE

Article Title: Osteocyte-Specific Deletion of Fgfr1 Suppresses FGF23

doi: 10.1371/journal.pone.0104154

Figure Lengend Snippet: (A) Gross appearance, tail length, and body weight. Compared with control mice, Fgfr1 Dmp1-cKO -null mice had normal gross appearance and body weight. However, Hyp mice showed considerably shorter tail length and lower body weight, compound Hyp ; Fgfr1 Dmp1-cKO -null mice displayed intermediate tail length and body weight between control and Hyp mice. Data are mean ± S.D. from 5–6 individual mice. (B and C) Western blot analysis of total Fgf2 and Fgf23 protein levels in bone. A representative Fgf2, Fgf23, and β-Actin gel were shown in upper, middle, and lower panels of B, respectively. The intensity of bands was quantified using Image J software ( http://rsb.info.nih.gov/ij/ ), and the data shown in C are mean ± S.D. from three independent experiments. Values sharing the same superscript in different groups are not significantly different at P <0.05.

Article Snippet: Antibody against Fgf23 (MAB2629) was obtained from R&D Systems, Inc. (Minneapolis, MN).

Techniques: Control, Western Blot, Software

Journal: PLoS ONE

Article Title: Osteocyte-Specific Deletion of Fgfr1 Suppresses FGF23

doi: 10.1371/journal.pone.0104154

Figure Lengend Snippet: Gene-expression profiles in bone in 6-week-old mice.

Article Snippet: Antibody against Fgf23 (MAB2629) was obtained from R&D Systems, Inc. (Minneapolis, MN).

Techniques:

Journal: PLoS ONE

Article Title: Osteocyte-Specific Deletion of Fgfr1 Suppresses FGF23

doi: 10.1371/journal.pone.0104154

Figure Lengend Snippet: Biochemistry analysis in 6-week-old mice.

Article Snippet: Antibody against Fgf23 (MAB2629) was obtained from R&D Systems, Inc. (Minneapolis, MN).

Techniques:

Journal: PLoS ONE

Article Title: Osteocyte-Specific Deletion of Fgfr1 Suppresses FGF23

doi: 10.1371/journal.pone.0104154

Figure Lengend Snippet: MC3T3-E1 cells were co-transfected with various plasmids and treated with different drugs as described in Material and Methods. (A) Dose-dependent stimulation of mouse FGF23 promoter activity by recombinant FGF2 (5∼100 ng/ml); (B) Dominate-negative FGFR1(TK−) constructs blocked the stimulatory effect of recombinant FGF2 (50 ng/ml) on mouse FGF23 promoter activity; (C) Effect of PLCγ inhibitor (U73122) on FGF2-induced FGF23 promoter activity; (D) Effect of PI3K inhibitor Wortmannin (Wort) on FGF2-induced FGF23 promoter activity; (E) Dose-dependent inhibition of FGF2-induced FGF23 promoter activation by MAPK inhibitors (U0126). (F) Overexpression of HMW- Fgf2 constructs stimulated mouse FGF23 promoter activity. Data are expressed as the mean ± S.D. from triple three independent experiments. Values sharing the same superscript in different groups are not significantly different at P <0.05. * indicates significant difference from control vector group.

Article Snippet: Antibody against Fgf23 (MAB2629) was obtained from R&D Systems, Inc. (Minneapolis, MN).

Techniques: Transfection, Activity Assay, Recombinant, Construct, Inhibition, Activation Assay, Over Expression, Control, Plasmid Preparation

Journal: PLoS ONE

Article Title: Osteocyte-Specific Deletion of Fgfr1 Suppresses FGF23

doi: 10.1371/journal.pone.0104154

Figure Lengend Snippet: MC3T3-E1 cells were co-transfected with pcDNA3.1- FGFR1 expression plasmids and human pcDNA3.1-FGF23-V5-His expression plasmids and treated with different drugs as described in Material and Methods. (A and B) Time-dependent stimulation of human FGF23-V5 protein expression by recombinant FGF2 (50 ng/ml); (C and D) Effect of PI3K inhibitor Wortmannin (Wort) and protein synthesis inhibitor Cycloheximide (CXM) on FGF2-induced human FGF23-V5 protein expression. Both Wortmannin and Cycloheximide completely blocked FGF2-induced FGF23 protein increase. Data are expressed as the mean ± S.D. from triple three independent experiments. Values sharing the same superscript in different groups are not significantly different at P <0.05.

Article Snippet: Antibody against Fgf23 (MAB2629) was obtained from R&D Systems, Inc. (Minneapolis, MN).

Techniques: Transfection, Expressing, Recombinant