Journal: Kidney & Blood Pressure Research

Article Title: Thromboxane A2 or Activated Platelets Slightly Lower Fgf23 Expression in vitro

doi: 10.1159/000545696

Figure Lengend Snippet: Fgf23 expression is down-regulated by pTxA2 in UMR-106 and MC3T3-E1 cells. Arithmetic means ± SEM of rel. Fgf23 gene expression in UMR-106 ( a , n = 5) and MC3T3-E1 ( b , n = 5) cells treated without or with pTxA2 for 24 h ( a ) or 6 h ( b ). * p < 0.05. SEM, standard error of the mean.

Article Snippet: The FGF23 protein concentration was determined using a mouse/rat C-terminal and a mouse/rat intact FGF23 ELISA kit (both from Quidel, San Clemente, CA, USA).

Techniques: Expressing, Gene Expression

Journal: Kidney & Blood Pressure Research

Article Title: Thromboxane A2 or Activated Platelets Slightly Lower Fgf23 Expression in vitro

doi: 10.1159/000545696

Figure Lengend Snippet: Fgf23 expression is decreased by TxA2 agonist I-BOP in UMR-106 and MC3T3-E1 cells. Arithmetic means ± SEM of rel. Fgf23 gene expression in UMR-106 ( a , n = 6) and in MC3T3-E1 ( b , n = 7) cells treated without or with TxA2 agonist I-BOP for 24 h ( a ) or 6 h ( b ). * p < 0.05. SEM, standard error of the mean.

Article Snippet: The FGF23 protein concentration was determined using a mouse/rat C-terminal and a mouse/rat intact FGF23 ELISA kit (both from Quidel, San Clemente, CA, USA).

Techniques: Expressing, Gene Expression

Journal: Kidney & Blood Pressure Research

Article Title: Thromboxane A2 or Activated Platelets Slightly Lower Fgf23 Expression in vitro

doi: 10.1159/000545696

Figure Lengend Snippet: Fgf23 expression is inhibited by TxA2 agonist U46619 in UMR-106 and MC3T3-E1 cells. Arithmetic means ± SEM ( n = 5) of rel. Fgf23 gene expression in UMR-106 ( a ) and MC3T3-E1 ( b ) cells treated without or with TxA2 agonist U46619 for 24 h ( a ) or 6 h ( b ). * p < 0.05. SEM, standard error of the mean.

Article Snippet: The FGF23 protein concentration was determined using a mouse/rat C-terminal and a mouse/rat intact FGF23 ELISA kit (both from Quidel, San Clemente, CA, USA).

Techniques: Expressing, Gene Expression

Journal: Kidney & Blood Pressure Research

Article Title: Thromboxane A2 or Activated Platelets Slightly Lower Fgf23 Expression in vitro

doi: 10.1159/000545696

Figure Lengend Snippet: C-terminal FGF23 concentration in the supernatant of UMR-106 cells is lowered by TxA2 agonists. Arithmetic means ± SEM of C-terminal FGF23 concentration in the supernatant of UMR-106 cells treated without or with 100 ng/mL pTxA2 ( a , n = 7), 100 ng/mL I-BOP ( b , n = 7) or 5 µ m U46619 ( c , n = 8) for 24 h. * p < 0.05, ** p < 0.01. SEM, standard error of the mean.

Article Snippet: The FGF23 protein concentration was determined using a mouse/rat C-terminal and a mouse/rat intact FGF23 ELISA kit (both from Quidel, San Clemente, CA, USA).

Techniques: Concentration Assay

Journal: Kidney & Blood Pressure Research

Article Title: Thromboxane A2 or Activated Platelets Slightly Lower Fgf23 Expression in vitro

doi: 10.1159/000545696

Figure Lengend Snippet: TxA2 antagonist SQ29548 reverses the effect of I-BOP on Fgf23 mRNA transcripts. Arithmetic means ± SEM ( n = 10) of relative Fgf23 gene expression in UMR-106 cells treated without or with 100 ng/mL TxA2 agonist I-BOP in the presence or absence of 1 µ m TxA2 antagonist SQ29548 for 24 h. ** p < 0.01. SEM, standard error of the mean.

Article Snippet: The FGF23 protein concentration was determined using a mouse/rat C-terminal and a mouse/rat intact FGF23 ELISA kit (both from Quidel, San Clemente, CA, USA).

Techniques: Gene Expression

Journal: Kidney & Blood Pressure Research

Article Title: Thromboxane A2 or Activated Platelets Slightly Lower Fgf23 Expression in vitro

doi: 10.1159/000545696

Figure Lengend Snippet: Exposure to activated platelets suppresses Fgf23 mRNA transcripts in UMR-106 cells. Arithmetic means ± SEM ( n = 8) of relative Fgf23 gene expression in UMR-106 cells exposed or not exposed to 100,000/µL freshly isolated human platelets in the presence or absence of 1 U/mL thrombin. * p < 0.05. SEM, standard error of the mean.

Article Snippet: The FGF23 protein concentration was determined using a mouse/rat C-terminal and a mouse/rat intact FGF23 ELISA kit (both from Quidel, San Clemente, CA, USA).

Techniques: Gene Expression, Isolation

Journal: PLOS ONE

Article Title: Phosphorus-independent role of FGF23 in erythropoiesis and iron homeostasis

doi: 10.1371/journal.pone.0315228

Figure Lengend Snippet: C57BL/6J male mice were fed a diet containing 0.6% inorganic phosphorus (Pi), 1.2% Pi, or 1.65% Pi for 2 weeks. (A) Body weight after 2 weeks of phosphorus supplementation. (B-C) Serum concentration of (B) intact and (C) C-terminal FGF23 measured by ELISA. (D-E) Phosphorus levels measured in (D) urine as ratio of phosphorus to creatinine and (E) serum. (F-H) Quantitative real-time RT-PCR for renal (F) α-Klotho , (G) Napi2a , and (H) Napi2c expression. Data are expressed as fold change (2 -ΔΔCt ) relative to housekeeping gene Hprt . Samples were measured in duplicates. Data are represented as mean ± SD. All data were analyzed for normality with Shapiro-Wilk test and homogeneity of variance by Brown-Forsythe test. For samples with normal distribution, one-way ANOVA was performed compared to 0.6% Pi with Dunnett’s multiple comparison test (B, C, E, F, G, H). When the samples were in normal distribution but not in homogeneity of variance, Welch’s ANOVA was performed (D). The samples not in normal distribution were analyzed with non-parametric Kruskal-Wallis test (A). (n = 7–10 per group). * P < 0.05, ** P < 0.01, *** P < 0.001 compared to 0.6% Pi.

Article Snippet: Serum FGF23 levels were measured using mouse FGF23 Intact and C-terminal ELISA assays (Quidel Corporation/Immutopics International, San Clemente, CA, USA).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Comparison

Journal: PLOS ONE

Article Title: Phosphorus-independent role of FGF23 in erythropoiesis and iron homeostasis

doi: 10.1371/journal.pone.0315228

Figure Lengend Snippet: C57BL/6J male mice were fed a diet containing 0.6% inorganic phosphorus (Pi), 1.2% Pi, or 1.65% Pi for 2 weeks. (A) Serum erythropoietin (EPO) levels measured by ELISA. (B-E) Quantitative real-time RT-PCR for expression of (B) renal Epo , (C) bone marrow Epo receptor (EpoR), (D) renal Hif-2α , and (E) bone marrow erythroferrone ( Erfe ). (F) Regression curve for correlation between bone marrow Erfe expression levels and serum FGF23 levels. (G) Quantitative real-time RT-PCR for splenic expression of heme oxygenase-1 ( Hmox-1 ). Data are expressed as fold change (2 -ΔΔCt ) relative to housekeeping gene Hprt . Data are represented as mean ± SD. All data were analyzed for normality with Shapiro-Wilk test and homogeneity of variance by Brown-Forsythe test. For samples with normal distribution and equal variance, one-way ANOVA with Dunnett’s multiple comparison test was performed (C, D, G). Samples with unequal variance were analyzed with Welch’s ANOVA (A, B, E). For the correlation curve, we applied simple linear and simple logistic regression (F). (n = 7–10 per group). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Serum FGF23 levels were measured using mouse FGF23 Intact and C-terminal ELISA assays (Quidel Corporation/Immutopics International, San Clemente, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Comparison

Journal: PLOS ONE

Article Title: Phosphorus-independent role of FGF23 in erythropoiesis and iron homeostasis

doi: 10.1371/journal.pone.0315228

Figure Lengend Snippet: Eight-week old C57BL/6J male mice were fed a diet containing 0.02% inorganic phosphorus (LP) for 2 weeks and compared to C57BL/6J male mice fed normal phosphorus diet (0.6% Pi; CONT). (A-B) Serum concentration of (A) intact FGF23, and (B) C-terminal FGF23 after 2 weeks of phosphorus restriction. (C-E) Circulating red blood cell parameters. (C) Red Blood Cells (RBCs), (D) Hemoglobin (Hgb), (E) Hematocrit (Hct). (F) Colony forming unit assay. Bone marrow cells were isolated from femora and tibiae, cultured for 12 days on methylcellulose medium, and counted for erythroid progenitor cells, burst forming unit-erythroid (BFU-E). (G) Quantitative real-time RT-PCR for renal Epo expression. Data are expressed as fold change (2 -ΔΔCt ) relative to housekeeping gene Hprt . (H) Serum concentration of EPO. Samples were measured in duplicates. (I) Quantitative real-time RT-PCR for splenic Hmox expression. Data are expressed as fold change (2 -ΔΔCt ) relative to housekeeping gene Hprt . Data are represented as mean ± SD. (n = 7–10 per group for A, B, C, D, E, F, H, and I; n = 5–6 per group for G). All data were analyzed for normality with Shapiro-Wilk test and homogeneity of variance by F test. For samples with normal distribution, unpaired t test was performed compared to CONT (B, C, D, E, F, G, H). When the samples were in normal distribution but not in homogeneity of variance, the data were analyzed by Welch’s t test (A, I). * P < 0.05, ** P < 0.01, *** P < 0.001 compared to CONT (control diet).

Article Snippet: Serum FGF23 levels were measured using mouse FGF23 Intact and C-terminal ELISA assays (Quidel Corporation/Immutopics International, San Clemente, CA, USA).

Techniques: Concentration Assay, Colony-forming Unit Assay, Isolation, Cell Culture, Quantitative RT-PCR, Expressing, Control

Journal: PLOS ONE

Article Title: Phosphorus-independent role of FGF23 in erythropoiesis and iron homeostasis

doi: 10.1371/journal.pone.0315228

Figure Lengend Snippet: Analysis of hematological and iron parameters in eight-week-old male mice carrying a mutation in the Phex gene (hyp mice) compared to age and sex matched wild-type littermates (control). (A-C) Serum concentration of (A) Phosphorus , (B) intact FGF23, and (C) C-terminal FGF23. (D-F) Circulating red blood cell parameters. (D) Red Blood Cells (RBCs), (E) Hemoglobin (Hgb), (F) Hematocrit (Hct). (G-H) Flow cytometry analysis of bone marrow erythroid progenitor cells from hyp and wild-type (control) mice. Percent of (G) pro-erythroblasts (pro-E) stained positive for Ter119 med and CD71 high and (H) terminally differentiated erythroid cells stained positive for Ter119 high and negative for CD71. (I) Colony forming unit assay. Bone marrow cells were isolated from femora and tibiae, cultured for 12 days on methylcellulose medium, and counted for erythroid progenitor cells, burst forming unit-erythroid (BFU-E). (J-L) Serum concentration of (J) EPO and (K) iron. (L) Liver iron content was measured by the ferrozine colorimetric assay and normalized to weight (mg) of dried tissue sample. Tissues were weighed at collection and their weight was normalized to total body weight of the animal and represented as percentage. Samples were measured in duplicates. Data are represented as mean ± SD. (n = 5–8 per group). All data were analyzed for normality with Shapiro-Wilk test and homogeneity of variance by F test. For samples with normal distribution, unpaired t test was performed compared to WT (B, C, D, E, G, H, I, J, K). When the samples were in normal distribution but not in homogeneity of variance, the data were analyzed by Welch’s t test (F, L). When samples were not normally distributed for parametric analysis, the data were analyzed by a non-parametric Mann-Whitney test (A). * P < 0.05, ** P < 0.01, *** P < 0.001 compared to wild-type (WT).

Article Snippet: Serum FGF23 levels were measured using mouse FGF23 Intact and C-terminal ELISA assays (Quidel Corporation/Immutopics International, San Clemente, CA, USA).

Techniques: Mutagenesis, Control, Concentration Assay, Flow Cytometry, Staining, Colony-forming Unit Assay, Isolation, Cell Culture, Colorimetric Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Transgenic augmentation of erythroferrone in mice ameliorates anemia in adenine-induced chronic kidney disease

doi: 10.1101/2024.12.06.627111

Figure Lengend Snippet: WT and TG mice from (n=6-7 mice/group) were analyzed. Measured parameters include ( A ) serum phosphate, ( B ) serum intact iFGF23, and qPCR analysis of kidney tissue for ( C-G ) Slc34a1 and Slc34a3 ( encoding sodium-dependent phosphate transporters 2A and 2C), Cyp27b1 and Cyp24a1 ( encoding the enzymes that respectively generate and break down the active form of vitamin D, 1,25-dihydroxyvitamin D3) and Kl (encoding Klotho, the renal coreceptor for iFGF23). Data are mean ± SEM, analyzed by unpaired- t -test with Welch’s correction (two-tailed). *P <.05; ns = non-significant.

Article Snippet: Intact bioactive FGF23 (iFGF23) levels were assessed by ELISA (60-6800, QuidelOrtho).

Techniques: Two Tailed Test