fgf23 Search Results


recombinant mouse fgf 23  (R&D Systems)
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R&D Systems recombinant mouse fgf 23
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rat anti mouse fgf23  (R&D Systems)
99
R&D Systems rat anti mouse fgf23
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fgf23  (R&D Systems)
94
R&D Systems fgf23
Figure 1. A novel kidney-specific protein regulates Pi homeostasis. siRNA library screening identified that the TMEM174 gene regulates Pi uptake in cells expressing NPT2A. (A) siRNA knockdown efficiency, (B) total Pi uptake, and (C) levels of NPT2A protein. Human renal tubular cells expressing NPT2A were treated with 20 nM siRNA and then with <t>FGF23</t> (10 nM) in the presence of 100 ng/ml secretory KL and 2 mg/ml heparin for 1 hour. (D) TMEM174 and NPT2A protein expression in human proximal tubular cells treated with 20 nM TMEM174 siRNA and then treated with FGF23 (10 nM) for 1–4 hours in the presence of 100 ng/ml secretory KL, 2 mg/ml heparin, and 0.1% BSA. Flarebio and Novus TMEM174 antibodies were used for human proximal tubular cells. The asterisk (*) represents nonspecific protein signal. (E) TMEM174 and NPT2A protein expression in OK-P cells treated with 20 nM TMEM174 siRNA and then with PTH (10 nM) for 1–4 hours in the presence of 0.1% BSA. Biomatik TMEM174 antibody was used for OK-P cells. Statistical analysis was performed using a one-way ANOVA. The experiments were repeated three times. ***P,0.001, **P,0.01 (ver- sus Scr1FGF23), #P,0.05 (versus Scr-FGF23).
Fgf23, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rhfgf23  (R&D Systems)
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R&D Systems rhfgf23
Figure 1. A novel kidney-specific protein regulates Pi homeostasis. siRNA library screening identified that the TMEM174 gene regulates Pi uptake in cells expressing NPT2A. (A) siRNA knockdown efficiency, (B) total Pi uptake, and (C) levels of NPT2A protein. Human renal tubular cells expressing NPT2A were treated with 20 nM siRNA and then with <t>FGF23</t> (10 nM) in the presence of 100 ng/ml secretory KL and 2 mg/ml heparin for 1 hour. (D) TMEM174 and NPT2A protein expression in human proximal tubular cells treated with 20 nM TMEM174 siRNA and then treated with FGF23 (10 nM) for 1–4 hours in the presence of 100 ng/ml secretory KL, 2 mg/ml heparin, and 0.1% BSA. Flarebio and Novus TMEM174 antibodies were used for human proximal tubular cells. The asterisk (*) represents nonspecific protein signal. (E) TMEM174 and NPT2A protein expression in OK-P cells treated with 20 nM TMEM174 siRNA and then with PTH (10 nM) for 1–4 hours in the presence of 0.1% BSA. Biomatik TMEM174 antibody was used for OK-P cells. Statistical analysis was performed using a one-way ANOVA. The experiments were repeated three times. ***P,0.001, **P,0.01 (ver- sus Scr1FGF23), #P,0.05 (versus Scr-FGF23).
Rhfgf23, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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intact fgf 23  (R&D Systems)
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R&D Systems intact fgf 23
Figure 1. A novel kidney-specific protein regulates Pi homeostasis. siRNA library screening identified that the TMEM174 gene regulates Pi uptake in cells expressing NPT2A. (A) siRNA knockdown efficiency, (B) total Pi uptake, and (C) levels of NPT2A protein. Human renal tubular cells expressing NPT2A were treated with 20 nM siRNA and then with <t>FGF23</t> (10 nM) in the presence of 100 ng/ml secretory KL and 2 mg/ml heparin for 1 hour. (D) TMEM174 and NPT2A protein expression in human proximal tubular cells treated with 20 nM TMEM174 siRNA and then treated with FGF23 (10 nM) for 1–4 hours in the presence of 100 ng/ml secretory KL, 2 mg/ml heparin, and 0.1% BSA. Flarebio and Novus TMEM174 antibodies were used for human proximal tubular cells. The asterisk (*) represents nonspecific protein signal. (E) TMEM174 and NPT2A protein expression in OK-P cells treated with 20 nM TMEM174 siRNA and then with PTH (10 nM) for 1–4 hours in the presence of 0.1% BSA. Biomatik TMEM174 antibody was used for OK-P cells. Statistical analysis was performed using a one-way ANOVA. The experiments were repeated three times. ***P,0.001, **P,0.01 (ver- sus Scr1FGF23), #P,0.05 (versus Scr-FGF23).
Intact Fgf 23, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse fgf23 elisa kit  (Elabscience Biotechnology)
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Elabscience Biotechnology mouse fgf23 elisa kit
Figure 1. Fucoidan Treatment Reversed the Changes of Serum Biochemical Indexes Caused by the Modeling. A–D, the Levels of BUN, Creatinine, ALP, and Pi in the Serum of Mice in the Five Groups were Detected by an Automatic Biochemical Analyzer. E–G, the Levels of iPTH, <t>FGF23,</t> and 1,25 (OH)2D3 in the Serum of Mice in the Five Groups Were Determined by ELISA. H, the Relative BMD of Mice in the Five Groups Was Measured by X-Ray Densitometers.
Mouse Fgf23 Elisa Kit, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human fgf23  (R&D Systems)
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R&D Systems human fgf23
Figure 1. Fucoidan Treatment Reversed the Changes of Serum Biochemical Indexes Caused by the Modeling. A–D, the Levels of BUN, Creatinine, ALP, and Pi in the Serum of Mice in the Five Groups were Detected by an Automatic Biochemical Analyzer. E–G, the Levels of iPTH, <t>FGF23,</t> and 1,25 (OH)2D3 in the Serum of Mice in the Five Groups Were Determined by ELISA. H, the Relative BMD of Mice in the Five Groups Was Measured by X-Ray Densitometers.
Human Fgf23, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fgf23 concentrations  (Cusabio)
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Cusabio fgf23 concentrations
Figure 1. Fucoidan Treatment Reversed the Changes of Serum Biochemical Indexes Caused by the Modeling. A–D, the Levels of BUN, Creatinine, ALP, and Pi in the Serum of Mice in the Five Groups were Detected by an Automatic Biochemical Analyzer. E–G, the Levels of iPTH, <t>FGF23,</t> and 1,25 (OH)2D3 in the Serum of Mice in the Five Groups Were Determined by ELISA. H, the Relative BMD of Mice in the Five Groups Was Measured by X-Ray Densitometers.
Fgf23 Concentrations, supplied by Cusabio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human fgf 23 elisa kit  (Elabscience Biotechnology)
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Elabscience Biotechnology human fgf 23 elisa kit
Figure 1. Fucoidan Treatment Reversed the Changes of Serum Biochemical Indexes Caused by the Modeling. A–D, the Levels of BUN, Creatinine, ALP, and Pi in the Serum of Mice in the Five Groups were Detected by an Automatic Biochemical Analyzer. E–G, the Levels of iPTH, <t>FGF23,</t> and 1,25 (OH)2D3 in the Serum of Mice in the Five Groups Were Determined by ELISA. H, the Relative BMD of Mice in the Five Groups Was Measured by X-Ray Densitometers.
Human Fgf 23 Elisa Kit, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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monoclonal anti rat fgf23 antibody  (R&D Systems)
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Fibroblast growth factor 23 increases mRNA levels of markers of hypertrophy and fibrosis. Rat cardiac myoblast cells (H9c2) were cultured with 0, 50, or 100 ng/mL fibroblast growth factor 3 <t>(FGF23)</t> for 24 h, and mRNA expression levels were analyzed by real-time PCR. (A) Atrial natriuretic factor (ANF) ( n = 6). ** P < 0.05 versus 0 ng/mL. (B) Brain natriuretic peptide (BNP) ( n = 6). * P < 0.01 versus 0 ng/mL. (C) β-myosin heavy chain ( beta MHC ) ( n = 6). ** P < 0.05 versus 0 ng/mL. (D) Alpha smooth muscle actin ( alpha SMA ) ( n = 6). * P < 0.01 versus 0 ng/mL. (E) Collagen I ( n = 6). * P < 0.01 versus 0 ng/mL. (F) FGF23 ( n = 6). * P < 0.01 versus 0 ng/mL. Data were analyzed by one-way analysis of variance.
Monoclonal Anti Rat Fgf23 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fgf23 e el r3031  (Elabscience Biotechnology)
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Elabscience Biotechnology fgf23 e el r3031
Fibroblast growth factor 23 increases mRNA levels of markers of hypertrophy and fibrosis. Rat cardiac myoblast cells (H9c2) were cultured with 0, 50, or 100 ng/mL fibroblast growth factor 3 <t>(FGF23)</t> for 24 h, and mRNA expression levels were analyzed by real-time PCR. (A) Atrial natriuretic factor (ANF) ( n = 6). ** P < 0.05 versus 0 ng/mL. (B) Brain natriuretic peptide (BNP) ( n = 6). * P < 0.01 versus 0 ng/mL. (C) β-myosin heavy chain ( beta MHC ) ( n = 6). ** P < 0.05 versus 0 ng/mL. (D) Alpha smooth muscle actin ( alpha SMA ) ( n = 6). * P < 0.01 versus 0 ng/mL. (E) Collagen I ( n = 6). * P < 0.01 versus 0 ng/mL. (F) FGF23 ( n = 6). * P < 0.01 versus 0 ng/mL. Data were analyzed by one-way analysis of variance.
Fgf23 E El R3031, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti fgf23 antibody  (R&D Systems)
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( A ) Description of the <t>FGF23-derived</t> transgenes (see Materials and Methods). ( B and C ) Western blot quantification of FGF23 isoform [cFGF23, FGF23, and FGF23-albumin (Alb) fusion] expression in the medium of Huh-7–transfected cells. (B) Representative Western blot. (C) Quantification of the FGF23 isoforms secreted in the medium of Huh-7 cells transfected with the indicated constructs. AU, arbitrary units. ( D to F ) C57BL6/J mice were injected with 1 × 10 12 vg per mouse of AAV8 vector expressing sp7-cFGF23co, sp7-cFGF23co-Alb, or sp7-cFGF23co-clFIX-Alb under the transcriptional control of the liver-specific hAAT promoter. (D) Representative Western blot of FGF23 isoforms measured in blood 1 month after vector injection. (E) Quantification of the expression of cFGF23-Alb fusion in blood. ( F ) Blood phosphate levels measured 1 month after vector injection. Statistical analyses were performed by analysis of variance (ANOVA) in (C) (** P < 0.01 versus intact FGF23; # P < 0.05 and ## P < 0.01 versus cFGF23 measured in cells transfected with native FGF23) and in (F) (* P < 0.05) and by Student’s t test in (E) (*** P < 0.001). All data are shown as means ± SD [ n = 3 independent transfections in (B and C); n = 4 to 5 in (E and F)].
Anti Fgf23 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Figure 1. A novel kidney-specific protein regulates Pi homeostasis. siRNA library screening identified that the TMEM174 gene regulates Pi uptake in cells expressing NPT2A. (A) siRNA knockdown efficiency, (B) total Pi uptake, and (C) levels of NPT2A protein. Human renal tubular cells expressing NPT2A were treated with 20 nM siRNA and then with FGF23 (10 nM) in the presence of 100 ng/ml secretory KL and 2 mg/ml heparin for 1 hour. (D) TMEM174 and NPT2A protein expression in human proximal tubular cells treated with 20 nM TMEM174 siRNA and then treated with FGF23 (10 nM) for 1–4 hours in the presence of 100 ng/ml secretory KL, 2 mg/ml heparin, and 0.1% BSA. Flarebio and Novus TMEM174 antibodies were used for human proximal tubular cells. The asterisk (*) represents nonspecific protein signal. (E) TMEM174 and NPT2A protein expression in OK-P cells treated with 20 nM TMEM174 siRNA and then with PTH (10 nM) for 1–4 hours in the presence of 0.1% BSA. Biomatik TMEM174 antibody was used for OK-P cells. Statistical analysis was performed using a one-way ANOVA. The experiments were repeated three times. ***P,0.001, **P,0.01 (ver- sus Scr1FGF23), #P,0.05 (versus Scr-FGF23).

Journal: Journal of the American Society of Nephrology

Article Title: Targeted Disruption of a Proximal Tubule–Specific TMEM174 Gene in Mice Causes Hyperphosphatemia and Vascular Calcification

doi: 10.1681/asn.2021121578

Figure Lengend Snippet: Figure 1. A novel kidney-specific protein regulates Pi homeostasis. siRNA library screening identified that the TMEM174 gene regulates Pi uptake in cells expressing NPT2A. (A) siRNA knockdown efficiency, (B) total Pi uptake, and (C) levels of NPT2A protein. Human renal tubular cells expressing NPT2A were treated with 20 nM siRNA and then with FGF23 (10 nM) in the presence of 100 ng/ml secretory KL and 2 mg/ml heparin for 1 hour. (D) TMEM174 and NPT2A protein expression in human proximal tubular cells treated with 20 nM TMEM174 siRNA and then treated with FGF23 (10 nM) for 1–4 hours in the presence of 100 ng/ml secretory KL, 2 mg/ml heparin, and 0.1% BSA. Flarebio and Novus TMEM174 antibodies were used for human proximal tubular cells. The asterisk (*) represents nonspecific protein signal. (E) TMEM174 and NPT2A protein expression in OK-P cells treated with 20 nM TMEM174 siRNA and then with PTH (10 nM) for 1–4 hours in the presence of 0.1% BSA. Biomatik TMEM174 antibody was used for OK-P cells. Statistical analysis was performed using a one-way ANOVA. The experiments were repeated three times. ***P,0.001, **P,0.01 (ver- sus Scr1FGF23), #P,0.05 (versus Scr-FGF23).

Article Snippet: Primary renal proximal tubular cells (PCS400-010) were purchased from ATCC and were grown and maintained at 37 C, in an atmosphere of 5% carbon dioxide, in a growth media (PCS- 400-030) in the absence/presence of 10–100 nM FGF23 (2604-FG; R&D System) and 100 ng/ml of a secretory form of KL, which was synthesized and purified from HEK293- expressing pcDNA3.1-KL-His cells at the Cell Technologies Core Facility at the University of Colorado Denver.

Techniques: Library Screening, Expressing, Knockdown

Figure 3. Disruption of Pi homeostasis by TMEM174 deficiency. Serum (A) Pi, (B) FGF23, (C) PTH, and (D) 1,25(OH)2D in 8-week- old TMEM174 KO male mice fed a chow diet. (E) TMEM174, renal Pi cotransporters, and NHEFR1 in the renal BBM of TMEM174 KO mice. Flarebio TMEM174 antibody was used to detect mouse TMEM174. (F) Immunofluorescence analysis of NPT2A in the kidneys of TMEM174 KO mice.25–27 (G) Total Pi uptake and (H) sodium-independent Pi uptake in the renal BBM of TMEM174 KO mice. Urinary (I) Pi and (J) Ca excretion of TMEM174 KO mice. Urine was collected in metabolic cages for 24 hours. (K) Alizarin red staining, (L) calcified lesions, and (M) aortic Ca content in the aortic sinus of TMEM174 KO mice. Statistical analysis was performed using a two-tailed t test. *P,0.05, **P,0.01, ***P,0.001. WT, wild type.

Journal: Journal of the American Society of Nephrology

Article Title: Targeted Disruption of a Proximal Tubule–Specific TMEM174 Gene in Mice Causes Hyperphosphatemia and Vascular Calcification

doi: 10.1681/asn.2021121578

Figure Lengend Snippet: Figure 3. Disruption of Pi homeostasis by TMEM174 deficiency. Serum (A) Pi, (B) FGF23, (C) PTH, and (D) 1,25(OH)2D in 8-week- old TMEM174 KO male mice fed a chow diet. (E) TMEM174, renal Pi cotransporters, and NHEFR1 in the renal BBM of TMEM174 KO mice. Flarebio TMEM174 antibody was used to detect mouse TMEM174. (F) Immunofluorescence analysis of NPT2A in the kidneys of TMEM174 KO mice.25–27 (G) Total Pi uptake and (H) sodium-independent Pi uptake in the renal BBM of TMEM174 KO mice. Urinary (I) Pi and (J) Ca excretion of TMEM174 KO mice. Urine was collected in metabolic cages for 24 hours. (K) Alizarin red staining, (L) calcified lesions, and (M) aortic Ca content in the aortic sinus of TMEM174 KO mice. Statistical analysis was performed using a two-tailed t test. *P,0.05, **P,0.01, ***P,0.001. WT, wild type.

Article Snippet: Primary renal proximal tubular cells (PCS400-010) were purchased from ATCC and were grown and maintained at 37 C, in an atmosphere of 5% carbon dioxide, in a growth media (PCS- 400-030) in the absence/presence of 10–100 nM FGF23 (2604-FG; R&D System) and 100 ng/ml of a secretory form of KL, which was synthesized and purified from HEK293- expressing pcDNA3.1-KL-His cells at the Cell Technologies Core Facility at the University of Colorado Denver.

Techniques: Disruption, Staining, Two Tailed Test

Figure 1. Fucoidan Treatment Reversed the Changes of Serum Biochemical Indexes Caused by the Modeling. A–D, the Levels of BUN, Creatinine, ALP, and Pi in the Serum of Mice in the Five Groups were Detected by an Automatic Biochemical Analyzer. E–G, the Levels of iPTH, FGF23, and 1,25 (OH)2D3 in the Serum of Mice in the Five Groups Were Determined by ELISA. H, the Relative BMD of Mice in the Five Groups Was Measured by X-Ray Densitometers.

Journal: Pharmacognosy Magazine

Article Title: Fucoidan Attenuated Kidney and Bone Damage Caused by CKD-MBD in Mice by Upregulating Klotho

doi: 10.1177/09731296231172549

Figure Lengend Snippet: Figure 1. Fucoidan Treatment Reversed the Changes of Serum Biochemical Indexes Caused by the Modeling. A–D, the Levels of BUN, Creatinine, ALP, and Pi in the Serum of Mice in the Five Groups were Detected by an Automatic Biochemical Analyzer. E–G, the Levels of iPTH, FGF23, and 1,25 (OH)2D3 in the Serum of Mice in the Five Groups Were Determined by ELISA. H, the Relative BMD of Mice in the Five Groups Was Measured by X-Ray Densitometers.

Article Snippet: Enzyme-linked Immunosorbent Assay (ELISA) The contents of intact parathyroid hormone (iPTH), fibroblast growth factor 23 (FGF23), and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3, DHVD3) in the serum were measured by the Mouse iPTH ELISA Kit (E-EL-M0709, Elabscience, China), Mouse FGF23 ELISA Kit (E-EL-M2415C, Elabscience, China), and Mouse DHVD3 ELISA Kit (E-EL-0016C, Elabscience, China), respectively.

Techniques: Enzyme-linked Immunosorbent Assay

Fibroblast growth factor 23 increases mRNA levels of markers of hypertrophy and fibrosis. Rat cardiac myoblast cells (H9c2) were cultured with 0, 50, or 100 ng/mL fibroblast growth factor 3 (FGF23) for 24 h, and mRNA expression levels were analyzed by real-time PCR. (A) Atrial natriuretic factor (ANF) ( n = 6). ** P < 0.05 versus 0 ng/mL. (B) Brain natriuretic peptide (BNP) ( n = 6). * P < 0.01 versus 0 ng/mL. (C) β-myosin heavy chain ( beta MHC ) ( n = 6). ** P < 0.05 versus 0 ng/mL. (D) Alpha smooth muscle actin ( alpha SMA ) ( n = 6). * P < 0.01 versus 0 ng/mL. (E) Collagen I ( n = 6). * P < 0.01 versus 0 ng/mL. (F) FGF23 ( n = 6). * P < 0.01 versus 0 ng/mL. Data were analyzed by one-way analysis of variance.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: Fibroblast growth factor 23 increases mRNA levels of markers of hypertrophy and fibrosis. Rat cardiac myoblast cells (H9c2) were cultured with 0, 50, or 100 ng/mL fibroblast growth factor 3 (FGF23) for 24 h, and mRNA expression levels were analyzed by real-time PCR. (A) Atrial natriuretic factor (ANF) ( n = 6). ** P < 0.05 versus 0 ng/mL. (B) Brain natriuretic peptide (BNP) ( n = 6). * P < 0.01 versus 0 ng/mL. (C) β-myosin heavy chain ( beta MHC ) ( n = 6). ** P < 0.05 versus 0 ng/mL. (D) Alpha smooth muscle actin ( alpha SMA ) ( n = 6). * P < 0.01 versus 0 ng/mL. (E) Collagen I ( n = 6). * P < 0.01 versus 0 ng/mL. (F) FGF23 ( n = 6). * P < 0.01 versus 0 ng/mL. Data were analyzed by one-way analysis of variance.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

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

Indoxyl sulfate increases mRNA levels of markers of hypertrophy and fibrosis. H9c2 cells were cultured with 0, 0.25, or 1.0 mM indoxyl sulfate (IS) for 24 h, and mRNA expression levels were analyzed by real-time PCR. (A) Atrial natriuretic factor ( ANF ) ( n = 6). * P < 0.01 versus 0 mM. (B) Brain natriuretic peptide (BNP) ( n = 6). * P < 0.01 versus 0 mM. (C) β-myosin heavy chain ( beta MHC ) ( n = 6). ** P < 0.05 versus 0 mM. (D) Alpha smooth muscle actin ( alpha SMA ) ( n = 6). ** P < 0.05 versus 0 mM. (E) Collagen I ( n = 6). ** P < 0.05 versus 0 mM. (F) FGF23 ( n = 6). ** P < 0.05 versus 0 mM. Data were analyzed by one-way analysis of variance.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: Indoxyl sulfate increases mRNA levels of markers of hypertrophy and fibrosis. H9c2 cells were cultured with 0, 0.25, or 1.0 mM indoxyl sulfate (IS) for 24 h, and mRNA expression levels were analyzed by real-time PCR. (A) Atrial natriuretic factor ( ANF ) ( n = 6). * P < 0.01 versus 0 mM. (B) Brain natriuretic peptide (BNP) ( n = 6). * P < 0.01 versus 0 mM. (C) β-myosin heavy chain ( beta MHC ) ( n = 6). ** P < 0.05 versus 0 mM. (D) Alpha smooth muscle actin ( alpha SMA ) ( n = 6). ** P < 0.05 versus 0 mM. (E) Collagen I ( n = 6). ** P < 0.05 versus 0 mM. (F) FGF23 ( n = 6). ** P < 0.05 versus 0 mM. Data were analyzed by one-way analysis of variance.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

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

Indoxyl sulfate increases fibroblast growth factor 23 protein expression and fibroblast growth factor receptor 4 phosphorylation. (A,B) Western blotting of fibroblast growth factor 3 (FGF23) protein expression in H9c2 cells. H9c2 cells were cultured with 0, 0.25, or 1.0 mM indoxyl sulfate (IS) for 72 h. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 5). * P < 0.05 versus IS 0 mM. (C,D) Western blotting of fibroblast growth factor receptor 4 (FGFR4) protein expression and FGFR4 phosphorylation in H9c2 cells. H9c2 cells were cultured with 0, 0.25, or 1.0 mM IS for 72 h. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 5). * P < 0.05 versus IS 0 mM. (E–G) Western blotting of furin protein in H9c2 cells. H9c2 cells were cultured with 0 or 1.0 mM IS for 72 h. The images are from different parts of the same gel. Beta actin protein expression was examined as an internal control ( n = 6). Furin stained as two bands, furin precursor (96 KDa) and mature (90 KDa) forms. * P < 0.05 versus IS 0 mM. Data were analyzed by one-way analysis of variance.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: Indoxyl sulfate increases fibroblast growth factor 23 protein expression and fibroblast growth factor receptor 4 phosphorylation. (A,B) Western blotting of fibroblast growth factor 3 (FGF23) protein expression in H9c2 cells. H9c2 cells were cultured with 0, 0.25, or 1.0 mM indoxyl sulfate (IS) for 72 h. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 5). * P < 0.05 versus IS 0 mM. (C,D) Western blotting of fibroblast growth factor receptor 4 (FGFR4) protein expression and FGFR4 phosphorylation in H9c2 cells. H9c2 cells were cultured with 0, 0.25, or 1.0 mM IS for 72 h. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 5). * P < 0.05 versus IS 0 mM. (E–G) Western blotting of furin protein in H9c2 cells. H9c2 cells were cultured with 0 or 1.0 mM IS for 72 h. The images are from different parts of the same gel. Beta actin protein expression was examined as an internal control ( n = 6). Furin stained as two bands, furin precursor (96 KDa) and mature (90 KDa) forms. * P < 0.05 versus IS 0 mM. Data were analyzed by one-way analysis of variance.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Phospho-proteomics, Western Blot, Cell Culture, Control, Staining

Body weight, blood pressure, and laboratory data from animal experiments.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: Body weight, blood pressure, and laboratory data from animal experiments.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

Techniques:

(A,B) IS increases intact FGF23 expression in the heart. The expression of intact FGF23 in the heart was significantly increased after IS treatment. This effect was abrogated by FGFR4 inhibition as shown by Western blotting ( n = 3–6). Data were analyzed by one-way analysis of variance. * P < 0.05.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: (A,B) IS increases intact FGF23 expression in the heart. The expression of intact FGF23 in the heart was significantly increased after IS treatment. This effect was abrogated by FGFR4 inhibition as shown by Western blotting ( n = 3–6). Data were analyzed by one-way analysis of variance. * P < 0.05.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Inhibition, Western Blot

IS induces FGF23 via AhR in vitro . H9c2 cells were cultured with 0 or 1 mM IS for 6 h (for mRNA) or 48 h (for protein) after pretreatment with AhR siRNA or control siRNA (ctrl). (A) FGF23 mRNA levels were significantly increased by IS and downregulated with AhR siRNA ( n = 11). * P < 0.01, ** P < 0.05. (B,C) Western blotting of FGF23 protein expression in H9c2 cells. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 6). * P < 0.01, ** P < 0.05. (D,E) Western blotting of HIF1α protein expression in H9c2 cells. The images are different parts of the same gel. Alpha, beta-tubulin protein expression was used as an internal control ( n = 6). * P < 0.01, ** P < 0.05. (F) H9c2 cells were cultured with 0, 50, or 100 ng/mL FGF23 for 24 h, and mRNA expression levels were analyzed by real-time PCR. Levels of polypeptide N-acetylgalactosaminyltransferase 3 ( GALNT3 ) mRNA ( n = 6). * P < 0.01, ** P < 0.05. (G) H9c2 cells were cultured with 0, 0.25, or 1.0 mM IS for 24 h, and mRNA expression levels were analyzed by real-time PCR. Levels of GALNT3 mRNA ( n = 6). * P < 0.01, ** P < 0.05. (H) Levels of GALNT3 mRNA ( n = 9). * P < 0.01, ** P < 0.05. (I,J) Western blotting of GALNT3 protein expression in H9c2 cells. Alpha, beta-tubulin protein expression was used as an internal control ( n = 6). * P < 0.01, ** P < 0.05. Data were analyzed by one-way analysis of variance.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: IS induces FGF23 via AhR in vitro . H9c2 cells were cultured with 0 or 1 mM IS for 6 h (for mRNA) or 48 h (for protein) after pretreatment with AhR siRNA or control siRNA (ctrl). (A) FGF23 mRNA levels were significantly increased by IS and downregulated with AhR siRNA ( n = 11). * P < 0.01, ** P < 0.05. (B,C) Western blotting of FGF23 protein expression in H9c2 cells. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 6). * P < 0.01, ** P < 0.05. (D,E) Western blotting of HIF1α protein expression in H9c2 cells. The images are different parts of the same gel. Alpha, beta-tubulin protein expression was used as an internal control ( n = 6). * P < 0.01, ** P < 0.05. (F) H9c2 cells were cultured with 0, 50, or 100 ng/mL FGF23 for 24 h, and mRNA expression levels were analyzed by real-time PCR. Levels of polypeptide N-acetylgalactosaminyltransferase 3 ( GALNT3 ) mRNA ( n = 6). * P < 0.01, ** P < 0.05. (G) H9c2 cells were cultured with 0, 0.25, or 1.0 mM IS for 24 h, and mRNA expression levels were analyzed by real-time PCR. Levels of GALNT3 mRNA ( n = 6). * P < 0.01, ** P < 0.05. (H) Levels of GALNT3 mRNA ( n = 9). * P < 0.01, ** P < 0.05. (I,J) Western blotting of GALNT3 protein expression in H9c2 cells. Alpha, beta-tubulin protein expression was used as an internal control ( n = 6). * P < 0.01, ** P < 0.05. Data were analyzed by one-way analysis of variance.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

Techniques: In Vitro, Cell Culture, Control, Western Blot, Expressing, Real-time Polymerase Chain Reaction

IS increases hypertrophic markers via FGF23 in vitro . H9c2 cells were cultured with 0 or 1 mM IS for 24 h after pretreatment with FGF23 siRNA or control siRNA (ctrl), and mRNA expression levels were analyzed by real-time PCR. (A) ANF , (B) BNP , (C) beta MHC , (D) alpha SMA , (E) collagen I, and (F) FGF23 mRNA levels were significantly increased by IS and downregulated with FGF23 siRNA ( n = 6). Data were analyzed by one-way analysis of variance. * P < 0.01, ** P < 0.05.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: IS increases hypertrophic markers via FGF23 in vitro . H9c2 cells were cultured with 0 or 1 mM IS for 24 h after pretreatment with FGF23 siRNA or control siRNA (ctrl), and mRNA expression levels were analyzed by real-time PCR. (A) ANF , (B) BNP , (C) beta MHC , (D) alpha SMA , (E) collagen I, and (F) FGF23 mRNA levels were significantly increased by IS and downregulated with FGF23 siRNA ( n = 6). Data were analyzed by one-way analysis of variance. * P < 0.01, ** P < 0.05.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

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

IS increases FGFR4 via FGF23 in vitro . H9c2 cells were cultured with 0 or 1 mM IS for 48 h after pretreatment with FGF23 siRNA or control siRNA (ctrl). (A,B) Western blotting of FGF23 protein expression in H9c2 cells. (C,D) Western blotting of FGFR4 protein expression and FGFR4 phosphorylation in H9c2 cells. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 6). Data were analyzed by one-way analysis of variance. * P < 0.01, ** P < 0.05.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: IS increases FGFR4 via FGF23 in vitro . H9c2 cells were cultured with 0 or 1 mM IS for 48 h after pretreatment with FGF23 siRNA or control siRNA (ctrl). (A,B) Western blotting of FGF23 protein expression in H9c2 cells. (C,D) Western blotting of FGFR4 protein expression and FGFR4 phosphorylation in H9c2 cells. Alpha, beta-tubulin protein expression was examined as an internal control ( n = 6). Data were analyzed by one-way analysis of variance. * P < 0.01, ** P < 0.05.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

Techniques: In Vitro, Cell Culture, Control, Western Blot, Expressing, Phospho-proteomics

Potential mechanism by which IS may induce LVH via the FGF23-FGFR4 pathway. IS upregulates FGF23 mRNA via AhR and HIF1α. IS also increases GALNT3 gene expression, preventing furin-mediated degradation of intact FGF23. Increased FGF23 induces myocardial hypertrophy by FGFR4-dependent activation.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Indoxyl sulfate induces left ventricular hypertrophy via the AhR-FGF23-FGFR4 signaling pathway

doi: 10.3389/fcvm.2023.990422

Figure Lengend Snippet: Potential mechanism by which IS may induce LVH via the FGF23-FGFR4 pathway. IS upregulates FGF23 mRNA via AhR and HIF1α. IS also increases GALNT3 gene expression, preventing furin-mediated degradation of intact FGF23. Increased FGF23 induces myocardial hypertrophy by FGFR4-dependent activation.

Article Snippet: The following antibodies were used as primary antibodies: monoclonal anti-rat FGF23 antibody (1:500, MAB2629; R&D Systems); polyclonal anti-goat FGF23 antibody (1:1000, ab123502; Abcam, Cambridge, UK); polyclonal anti-rabbit FGFR4 antibody (1:1000, ab119378; Abcam); polyclonal anti-rabbit FGFR4 (phospho Y642; pFGFR4) antibody (1:1000, ab192589; Abcam); polyclonal anti-rabbit furin antibody (1:1000, PA1-062; Thermo Fisher Scientific); polyclonal anti-rabbit hypoxia-inducible factor 1 alpha (HIF1α) antibody (1:1000, NB100-134; Novus Biologicals, Centennial, CO); polyclonal anti-rabbit polypeptide GALNT3 antibody (1:1000, SAB2106736; Sigma-Aldrich); and anti-rabbit α/β tubulin (1:1000, CST#2148; Cell Signaling Technology, Danvers, MA).

Techniques: Gene Expression, Activation Assay

( A ) Description of the FGF23-derived transgenes (see Materials and Methods). ( B and C ) Western blot quantification of FGF23 isoform [cFGF23, FGF23, and FGF23-albumin (Alb) fusion] expression in the medium of Huh-7–transfected cells. (B) Representative Western blot. (C) Quantification of the FGF23 isoforms secreted in the medium of Huh-7 cells transfected with the indicated constructs. AU, arbitrary units. ( D to F ) C57BL6/J mice were injected with 1 × 10 12 vg per mouse of AAV8 vector expressing sp7-cFGF23co, sp7-cFGF23co-Alb, or sp7-cFGF23co-clFIX-Alb under the transcriptional control of the liver-specific hAAT promoter. (D) Representative Western blot of FGF23 isoforms measured in blood 1 month after vector injection. (E) Quantification of the expression of cFGF23-Alb fusion in blood. ( F ) Blood phosphate levels measured 1 month after vector injection. Statistical analyses were performed by analysis of variance (ANOVA) in (C) (** P < 0.01 versus intact FGF23; # P < 0.05 and ## P < 0.01 versus cFGF23 measured in cells transfected with native FGF23) and in (F) (* P < 0.05) and by Student’s t test in (E) (*** P < 0.001). All data are shown as means ± SD [ n = 3 independent transfections in (B and C); n = 4 to 5 in (E and F)].

Journal: Science Advances

Article Title: A novel therapeutic strategy for skeletal disorders: Proof of concept of gene therapy for X-linked hypophosphatemia

doi: 10.1126/sciadv.abj5018

Figure Lengend Snippet: ( A ) Description of the FGF23-derived transgenes (see Materials and Methods). ( B and C ) Western blot quantification of FGF23 isoform [cFGF23, FGF23, and FGF23-albumin (Alb) fusion] expression in the medium of Huh-7–transfected cells. (B) Representative Western blot. (C) Quantification of the FGF23 isoforms secreted in the medium of Huh-7 cells transfected with the indicated constructs. AU, arbitrary units. ( D to F ) C57BL6/J mice were injected with 1 × 10 12 vg per mouse of AAV8 vector expressing sp7-cFGF23co, sp7-cFGF23co-Alb, or sp7-cFGF23co-clFIX-Alb under the transcriptional control of the liver-specific hAAT promoter. (D) Representative Western blot of FGF23 isoforms measured in blood 1 month after vector injection. (E) Quantification of the expression of cFGF23-Alb fusion in blood. ( F ) Blood phosphate levels measured 1 month after vector injection. Statistical analyses were performed by analysis of variance (ANOVA) in (C) (** P < 0.01 versus intact FGF23; # P < 0.05 and ## P < 0.01 versus cFGF23 measured in cells transfected with native FGF23) and in (F) (* P < 0.05) and by Student’s t test in (E) (*** P < 0.001). All data are shown as means ± SD [ n = 3 independent transfections in (B and C); n = 4 to 5 in (E and F)].

Article Snippet: After transfer on nitrocellulose, the membrane was blocked and incubated with an anti-FGF23 antibody (R&D System, Minneapolis, MN).

Techniques: Derivative Assay, Western Blot, Expressing, Transfection, Construct, Injection, Plasmid Preparation, Control