recombinant human fgf23  (R&D Systems)

Journal: Cell Communication and Signaling : CCS

Article Title: The Roxadustat (FG-4592) ameliorates tubulointerstitial fibrosis by promoting intact FGF23 cleavage

doi: 10.1186/s12964-025-02175-2

Figure Lengend Snippet: FG-4592 lowers iFGF23 level in CKD rats and CKD patients. Parameters measured include ( A ) Serum intact fibroblast growth factor 23 (iFGF23) levels in CKD rats. B Serum total FGF23 levels in CKD rats. C Serum iFGF23 levels in CKD patients. D Serum total FGF23 levels in CKD patients. E Scatter plots with linear regression analysis demonstrating a significant correlation between blood urea nitrogen (BUN) and iFGF23 levels in CKD rats. F Scatter plots with linear regression analysis showing a significant correlation between Scr and iFGF23 levels in CKD rats. G Scatter plots with linear regression analysis revealing a significant correlation between iFGF23 levels and the percentage of fibrotic area in the kidney of CKD rats. H Scatter plots with linear regression analysis indicating a significant correlation between iFGF23 levels and Collagen 1 staining intensity in the kidney of CKD rats. I Scatter plots with linear regression analysis illustrating a significant correlation between iFGF23 levels and Fibronectin staining intensity in the kidney of CKD rats. J Scatter plots with linear regression analysis demonstrating a significant correlation between iFGF23 levels and α-SMA staining intensity in the kidney of CKD rats. Data are presented as means ± SD ( n = 5). Statistical significance is denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: CKD rats were surgically implanted with Alzet osmotic pumps (Durect Corporation, Cupertino, CA) as described previously [ ] and infused with recombinant FGF23 (rFGF23) (MCE, USA) or vehicle (normal saline; VE) for 28 days.

Techniques: Staining

Journal: Cell Communication and Signaling : CCS

Article Title: The Roxadustat (FG-4592) ameliorates tubulointerstitial fibrosis by promoting intact FGF23 cleavage

doi: 10.1186/s12964-025-02175-2

Figure Lengend Snippet: The renoprotective effects of FG-4592 were abrogated by FGF23 overexpression. Parameters measured include ( A ) Schematic diagram illustrating the experimental groups of rats: CKD + FG-4592 + vehicle and CKD + FG-4592 + rFGF23. B and C Renal function was assessed by measuring Scr and BUN levels. D mRNA expression levels of Col1a1 , Fn1 , and Acta2 in the renal cortex were quantified using real-time PCR. E Western blot analysis was performed to evaluate the protein expression of Collagen 1 and α-SMA, which reflect the extent of TIF. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) was used as the loading control. F Representative images of PAS staining and the percentage of fibrotic area in kidney tissues. Scale bars: 50 μm. G Representative immunohistochemical images demonstrating the upregulation of Collagen 1 and α-SMA in the CKD + FG-4592 + rFGF23 group. Scale bars: 50 μm. Data are presented as means ± SD ( n = 5). Statistical significance is denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: CKD rats were surgically implanted with Alzet osmotic pumps (Durect Corporation, Cupertino, CA) as described previously [ ] and infused with recombinant FGF23 (rFGF23) (MCE, USA) or vehicle (normal saline; VE) for 28 days.

Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Staining, Immunohistochemical staining

Journal: Cell Communication and Signaling : CCS

Article Title: The Roxadustat (FG-4592) ameliorates tubulointerstitial fibrosis by promoting intact FGF23 cleavage

doi: 10.1186/s12964-025-02175-2

Figure Lengend Snippet: iFGF23 mediates FG-4592-induced inhibition of tubular cell fibrogenesis. HK-2 cells were pre-treated with TGF-β1 (10 ng/mL) for 24 h, followed by treatment with recombinant FGF23 (rFGF23, 50 ng/mL) or vehicle for an additional 24 h. Parameters measured include: A COL1A1 mRNA expression levels in HK-2 cells, as determined by quantitative PCR (qPCR). B TGFB1 mRNA expression levels in HK-2 cells, as determined by qPCR. C Collagen 1 protein expression levels in HK-2 cells, as assessed by Western blot analysis. D Schematic illustration of the co-culture system using UMR-106 cells and HK-2 cells in a Transwell setup. E mRNA expression levels of Fn1 , Acta2 , and Col1a1 in HK-2 cells, as quantified by qPCR. Data are presented as means ± SD ( n = 5). Statistical significance is denoted as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

Article Snippet: CKD rats were surgically implanted with Alzet osmotic pumps (Durect Corporation, Cupertino, CA) as described previously [ ] and infused with recombinant FGF23 (rFGF23) (MCE, USA) or vehicle (normal saline; VE) for 28 days.

Techniques: Inhibition, Recombinant, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Co-Culture Assay

Journal: Cell Communication and Signaling : CCS

Article Title: The Roxadustat (FG-4592) ameliorates tubulointerstitial fibrosis by promoting intact FGF23 cleavage

doi: 10.1186/s12964-025-02175-2

Figure Lengend Snippet: FG-4592 ameliorates TIF by inhibiting iFGF23-WNT5A pathway. Parameters measured include ( A ) Hierarchical clustering analysis revealing distinct mRNA expression profiles among the experimental groups. Yellow and blue shades indicate expression levels above and below the relative mean, respectively, across all samples. B Gene Ontology (GO) enrichment analysis. C Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. D and F mRNA and protein expression levels of Wnt5a and β-catenin in the following groups: sham, CKD + vehicle, and CKD + FG-4592. E and G mRNA and protein expression levels of Wnt5a and β-catenin in the following groups: CKD + FG-4592 + vehicle and CKD + FG-4592 + rFGF23. H and I Representative immunohistochemical images showing Wnt5a and β-catenin expression in kidney tissues across different experimental groups. J mRNA expression levels of COL1A1 , ACTA1 , WNT5A , and CTNNB1 in HK-2 cells following WNT5A siRNA interference, as determined by quantitative PCR (qPCR). K Protein expression levels of Collagen 1, Wnt5a, and β-catenin in HK-2 cells after WNT5A siRNA interference, as assessed by Western blot analysis. Scale bars: 50 μm. Data are presented as means ± SD ( n = 5). Statistical significance is denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: CKD rats were surgically implanted with Alzet osmotic pumps (Durect Corporation, Cupertino, CA) as described previously [ ] and infused with recombinant FGF23 (rFGF23) (MCE, USA) or vehicle (normal saline; VE) for 28 days.

Techniques: Expressing, Immunohistochemical staining, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: The Roxadustat (FG-4592) ameliorates tubulointerstitial fibrosis by promoting intact FGF23 cleavage

doi: 10.1186/s12964-025-02175-2

Figure Lengend Snippet: FG-4592 lowers iFGF23 levels through Furin-mediated cleavage. Parameters measured include ( A ) mRNA expression levels of Furin , Galnt3 , and Fam20c in bone tissues from the following groups: sham, CKD + vehicle, and CKD + FG-4592, as determined by quantitative PCR (qPCR). B Protein levels of HIF-1α, Furin, and intact fibroblast growth factor 23 (iFGF23) in bone tissues treated with FG-4592 or vehicle, as assessed by Western blot analysis. C Representative images of hematoxylin and eosin (HE) staining of bone tissues (scale bars: 20 μm), along with IHC staining for HIF-1α and iFGF23 (scale bars: 50 μm) and Furin (scale bars: 100 μm). D UMR-106 cells were pretreated with 10 nM 1,25(OH) 2 D 3 , and the expression of FGF23 mRNA was analyzed using real-time PCR. E Western blot analysis was performed to measure iFGF23 protein expression in UMR-106 cells. Cells were pretreated with indoxyl sulfate (IS, 0.5 mM) or vehicle, followed by treatment with FG-4592 (50 μM) or vehicle for 24 h. F Levels of iFGF23 and total FGF23 in the cell supernatant were quantified using an ELISA assay kit. G Western blot analysis was used to evaluate the protein expression of iFGF23 and Furin in UMR-106 cells. H Motif map of the Furin promoter region, highlighting the HIF-1 binding site. I ChIP assays demonstrated HIF-1 binding to the Furin promoter, which was enhanced by FG-4592 stimulation (50 μM for 24 h). J Protein expression levels of Furin and iFGF23 in UMR-106 cells following Furin siRNA interference, as determined by Western blot analysis. K Levels of iFGF23 and total FGF23 in the cell supernatant were measured using an ELISA assay kit after Furin siRNA interference. Data are presented as means ± SD ( n = 5). Statistical significance is denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: CKD rats were surgically implanted with Alzet osmotic pumps (Durect Corporation, Cupertino, CA) as described previously [ ] and infused with recombinant FGF23 (rFGF23) (MCE, USA) or vehicle (normal saline; VE) for 28 days.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Staining, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Anti‐GBM disease causes tubular damage and partial renal resistance to FGF23. (A) Depicts the experimental workflow in male C57BL/6 mice undergoing induction of anti‐GBM disease using nephrotoxic serum followed by 6 days of intravenous (IV) FGF23 or vehicle treatment. (B) Shows the glomerular filtration rate of different experimental groups at days 0 and 7. (C) Shows urinary albumin/creatinine ratio at day 7. (D) Shows the example renal sections negative or positive for renal tubular casts (arrows) and quantitative tubular cast scores. (E) Shows plasma phosphate and fractional excretion of phosphate of healthy mice and mice with anti‐GBM treated with vehicle or FGF23. Analyses in panel (B): paired t ‐test. Analyses in panels (C–E): two‐way ANOVA. Anti‐GBM, anti‐glomerular basement membrane; d, disease state; t, treatment.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Filtration, Clinical Proteomics, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Six‐day course of FGF23 treatment induces renal transcriptional signatures of inflammatory responses and injury. (A) Indicates the number of differentially expressed genes according to experimental comparison in renal bulk RNA‐Seq. (B, C) Depict volcano plots of DEGs above a cutoff of adjusted p < 0.05 and log2‐fold change >1, in the comparison of FGF23 versus vehicle effect in mice with anti‐GBM (B) and the interaction between treatment and disease effect (C). (D–F) Depict significant Reactome gene set enrichment analyses of FGF23 effects in healthy mice (D), mice with anti‐GBM disease (E), and the interaction between treatment and disease effect (F). (G) Depicts a network of ligand–receptor interaction pairs that were significant for FGF23 versus vehicle comparisons in mice with anti‐GBM disease by bulk RNA‐Seq. The ligand–receptor interactions were inferred using R/BulkSignalR. Anti‐GBM, anti‐glomerular basement membrane disease. N = 3 for anti‐GBM groups and n = 4 for healthy groups.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Comparison, RNA Sequencing, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Bulk RNA‐Seq deconvolution and immunofluorescence staining reveal an FGF23‐driven increase in overall immune cell and macrophage abundance in the kidneys of mice with anti‐GBM disease. (A, B) Depict the annotation of renal cell clusters in the reanalysis of the single‐cell RNA‐Seq dataset GSE107585 of murine kidney from 7 sex‐mixed healthy C57BL/6 mice; see also Supporting Information Figure . (C) Shows a wedding pie plot of the bulk deconvolution of the renal cellular composition according to FGF23 treatment and anti‐GBM disease state, as indicated by labels. Overall renal immune cells and macrophage‐like cells are displayed by bulk deconvolution (D, G) and by immunofluorescence with automated quantification for CD45 (E, F) and F4/80 (H, I). RNA‐Seq: N = 3 for anti‐GBM groups and n = 4 for healthy groups. Immunofluorescence: n = 4 per group. Statistical analysis: two‐way ANOVA. anti‐GBM, anti‐glomerular basement membrane disease; Baso, basophil; CD, collecting duct; d, disease state; DCT, distal convoluted tubule; DLOH, descending limb of Henle; EC, endothelial cell; Granul, granulocyte; i, interaction; IC, intercalated cells; Ly, lymphocyte; Mono, monocyte; Mph, macrophage; NK, natural killer cell; PC, principal cells; PT, proximal tubule; S, segment; t, treatment.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: RNA Sequencing, Immunofluorescence, Staining, Single Cell, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Immune protein profiling highlights an increase in circulating soluble tumor necrosis factor receptors induced by FGF23 and anti‐GBM disease in mice. A plasma cytokine protein array shows FGF23 effects in healthy male C57BL/6 mice (A) and in mice treated with nephrotoxic serum to induce anti‐GBM disease (B). The interaction between treatment and disease state (C) and the overall disease effect (D) are shown. (E–F) Depict analyses of soluble TNF receptors 1 and 2 by two‐way ANOVA. d, disease state; t, treatment. N = 4 biologically independent replicates per group. Anti‐GBM, anti‐glomerular basement membrane.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Clinical Proteomics, Protein Array, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Renal immune cell recruitment driven by FGF23 excess is exposure time dependent. Renal microarray transcriptome datasets GDS3361 of male Fgf23 transgenic and control mice or sex‐matched Hyp mice of dataset GDS879 (B) underwent bulk deconvolution with reference to single‐cell RNA‐Seq dataset GSE107585 of murine kidney from 7 sex‐mixed healthy C57BL/6 mice and, subsequently, visualization of overall fractions of inferred immune cells and macrophage‐like cells, as indicated. (C) Shows the experimental workflow of experiments with female BALB/c mice undergoing Adriamycin (doxorubicin) nephropathy followed by a single intravenous (IV) injection of FGF23 or vehicle. (D) Shows the urinary albumin/creatinine ratio 7 days after induction of Adriamycin nephropathy. Statistical analysis: two‐way ANOVA. d, disease state. (E–G) Show significant Reactome gene set enrichment analyses of renal FGF23 effects in healthy mice (E), mice with Adriamycin nephropathy (F), and the interaction between treatment and disease effect (G). (H) Depicts the log‐fold change of 8 transcripts with lowest adjusted p ‐value in the interaction analysis of FGF23 effect in diseased versus FGF23 effect in healthy mice in a 2 × 2 factorial design. N = 5 (A), 10 (B), or 4 (C–H) biologically independent replicates per group.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Microarray, Transgenic Assay, Control, Single Cell, RNA Sequencing, IV Injection

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Intrarenal proinflammatory effects of FGF23 applied ex vivo in PCKS. (A) Male DBA/2J mice underwent dietary treatment with 0.2% adenine for 15 weeks, followed by organ collection and preparation of 300 µm PCKS for a 24‐h treatment with FGF23 or vehicle ex vivo. No disease‐free controls were used for this substudy. (B) Depicts the fibrotic changes in a representative 4 µm section of PCKS stained with hematoxylin and eosin (left scale bar, 500 µm; right, 100 µm). (C) Shows the log‐fold change of upregulated transcripts with the lowest adjusted p ‐value in the FGF23 versus vehicle comparison. (D–F) Show gene set enrichment analyses of FGF23 effects in Reactome (D), WikiPathways (E), and Pathway Interaction Database (F) gene sets. N = 4 biologically independent replicates per group. PCKS, precision‐cut kidney slices.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Ex Vivo, Staining, Comparison

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: FGF23 is associated with renal immune cell content in human patients with IgA nephropathy. As a reference, single‐nucleus RNA‐Seq data from 5 human kidney biopsies ( GSE199711 ) of 2 healthy controls and 3 patients with chronic kidney disease (CKD) were reanalyzed and annotated (A); see also Supporting Information Figure . 35 patients with IgA nephropathy from the Karolinska Kidney Biopsy Cohort showed an inverse univariable association between circulating FGF23 and measured glomerular filtration rate (GFR) (B). The associations between transcriptome‐inferred renal fibroblasts and immune cells (C–D) or macrophages (E–F) and circulating FGF23 are shown, with adjustment for GFR, 25OH‐vitamin D and parathyroid hormone. (D, F) Show disaggregated data stratified by GFR in CKD stages I–II and III–V.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: RNA Sequencing, Filtration