Journal: BMC Medicine

Article Title: The anti-aging factor Klotho protects against acquired long QT syndrome induced by uremia and promoted by fibroblast growth factor 23

doi: 10.1186/s12916-021-02209-9

Figure Lengend Snippet: Demographic data of patients with dialysis-dependent CKD by quartile of circulating FGF23 levels

Article Snippet: Adult 14-week-old male C57BL/6 J mice (Charles River Laboratories International Inc.) were used to study the effect of FGF23 on heart rhythm by a single-dose intraperitoneal injection of recombinant mouse FGF23 (40 μg/kg) (R&D Systems) during ECG recording.

Techniques:

Journal: BMC Medicine

Article Title: The anti-aging factor Klotho protects against acquired long QT syndrome induced by uremia and promoted by fibroblast growth factor 23

doi: 10.1186/s12916-021-02209-9

Figure Lengend Snippet: FGF23 induces prolonged ventricular repolarization in healthy mice. A ECG waveforms in wild-type healthy mice before (black line) and after (red line) FGF23 administration (40 μg/kg). B–F Time course of QRS interval ( B ), QT interval ( C ), QTc interval ( D ), JT interval ( E ), and T peak to T end (TpTe) interval ( F ) measures from ECG recordings after vehicle ( N = 5 mice) or FGF23 ( N = 9 mice) administration. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001 versus baseline

Article Snippet: Adult 14-week-old male C57BL/6 J mice (Charles River Laboratories International Inc.) were used to study the effect of FGF23 on heart rhythm by a single-dose intraperitoneal injection of recombinant mouse FGF23 (40 μg/kg) (R&D Systems) during ECG recording.

Techniques:

Journal: BMC Medicine

Article Title: The anti-aging factor Klotho protects against acquired long QT syndrome induced by uremia and promoted by fibroblast growth factor 23

doi: 10.1186/s12916-021-02209-9

Figure Lengend Snippet: Macroscopic and biochemical parameters in the experimental CKD model and after treatment with recombinant Klotho

Article Snippet: Adult 14-week-old male C57BL/6 J mice (Charles River Laboratories International Inc.) were used to study the effect of FGF23 on heart rhythm by a single-dose intraperitoneal injection of recombinant mouse FGF23 (40 μg/kg) (R&D Systems) during ECG recording.

Techniques: Recombinant

Journal: BMC Medicine

Article Title: The anti-aging factor Klotho protects against acquired long QT syndrome induced by uremia and promoted by fibroblast growth factor 23

doi: 10.1186/s12916-021-02209-9

Figure Lengend Snippet: Klotho prevents Kv4.2 α-subunit downregulation in ventricular myocytes from Nfx and after FGF23 incubation. A mRNA expression of Kv4.2 channel subunit in Sham ( N = 9 mice), Nfx ( N = 8 mice), Sham+rKL ( N = 9 mice), and Nfx+rKL ( N = 7 mice) mice. B mRNA expression of Kv4.2 in neonatal ventricular cardiomyocytes incubated with vehicle ( N = 6 replicate wells), FGF23 ( N = 9 replicate wells), rKL ( N = 9 replicate wells), and FGF23 + rKL ( N = 11 replicate wells). Data are presented as mean ± SEM. * P < 0.05 vs. the corresponding control (Sham or vehicle-neonatal cardiomyocytes) and # P < 0.05 vs. FGF23-neonatal cardiomyocytes

Article Snippet: Adult 14-week-old male C57BL/6 J mice (Charles River Laboratories International Inc.) were used to study the effect of FGF23 on heart rhythm by a single-dose intraperitoneal injection of recombinant mouse FGF23 (40 μg/kg) (R&D Systems) during ECG recording.

Techniques: Incubation, Expressing, Control

Journal: BMC Medicine

Article Title: The anti-aging factor Klotho protects against acquired long QT syndrome induced by uremia and promoted by fibroblast growth factor 23

doi: 10.1186/s12916-021-02209-9

Figure Lengend Snippet: Diagram representing the involvement of FGF23-Klotho axis in acquired long QT syndrome. CKD, chronic kidney disease; FGF23, fibroblast growth factor 23; I tof , fast transient outward potassium current; kl/kl , Klotho hypomorphic mice; Kv4.2, K + channel 4.2 subunit; Nfx, nephrectomy; sKL, soluble Klotho; Tg-kl, Klotho overexpression mice

Article Snippet: Adult 14-week-old male C57BL/6 J mice (Charles River Laboratories International Inc.) were used to study the effect of FGF23 on heart rhythm by a single-dose intraperitoneal injection of recombinant mouse FGF23 (40 μg/kg) (R&D Systems) during ECG recording.

Techniques: Over Expression

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: eLife

Article Title: High-phytate/low-calcium diet is a risk factor for crystal nephropathies, renal phosphate wasting, and bone loss

doi: 10.7554/eLife.52709

Figure Lengend Snippet: Time-course analysis of serum levels of intact FGF23 for three consecutive phytate doses. Statistical significance was tested by repeated measure ANOVA between subjects and groups. All data are presented as the mean ± SD for each dose (n = 6 per dose).

Article Snippet: Rat intact PTH (Immutopics, San Clemente, CA, USA), human intact PTH (Abcam), soluble RANKL (Immundiagnostik, Bensheim, Germany), osteoprotegerin (Alpco Immunoassay, Salem, NH, USA), rat intact and C-terminal FGF23 (Elabscience, Houston, TX, USA), and human intact FGF23 (Elabscience) were measured using enzyme-linked immunosorbent assay.

Techniques:

Journal: eLife

Article Title: High-phytate/low-calcium diet is a risk factor for crystal nephropathies, renal phosphate wasting, and bone loss

doi: 10.7554/eLife.52709

Figure Lengend Snippet: ( A–C ) Time-course analysis of serum levels of intact PTH ( A ), 25(OH)D ( B ), and 1,25(OH) 2 D ( C ) in rats fed control, HP-LCa 2+ , and HP-HCa 2+ diets. For the early measurement of 25(OH)D ( B ) and 1,25(OH) 2 D, we pooled the sera from 2 to 3 rats. ( D–F ) Time-course analysis of renal CYP27B1 ( D ), CYP24A1 ( E ), and VDR ( F ) in rats fed control, HP-LCa 2+ , and HP-HCa 2+ diets. ( G ) Immunoblot analysis of renal αKlotho, NHERF1, NaPi-2a in rats fed control, HP-LCa 2+ , and HP-HCa 2+ diets. ( H ) The levels of renal proteins were quantified using ImageJ software (NIH, Bethesda, MD, USA). ( I ) Immunoblot analysis of calvarial FGF23 in rats fed control, HP-LCa 2+ , and HP-HCa 2+ diets. ( J ) The levels of total calvarial FGF23 were quantified using ImageJ software. All comparisons were conducted using two-way ANOVA with Tukey’s post hoc multiple comparison testing or the Kruskal–Wallis test ( A–C ) for non-parametric statistical analysis of data that did not have a normal distribution. Panels ( A–C ) failed the normality test, but no outliers were identified. All data are presented as the mean ± SD of each group (n = 4–6 per group). *, p<0.05; **, p<0.01; ***, p<0.001, compared with controls.

Article Snippet: Rat intact PTH (Immutopics, San Clemente, CA, USA), human intact PTH (Abcam), soluble RANKL (Immundiagnostik, Bensheim, Germany), osteoprotegerin (Alpco Immunoassay, Salem, NH, USA), rat intact and C-terminal FGF23 (Elabscience, Houston, TX, USA), and human intact FGF23 (Elabscience) were measured using enzyme-linked immunosorbent assay.

Techniques: Control, Western Blot, Software, Comparison

Journal: eLife

Article Title: High-phytate/low-calcium diet is a risk factor for crystal nephropathies, renal phosphate wasting, and bone loss

doi: 10.7554/eLife.52709

Figure Lengend Snippet: ( A ) The renal expression of downregulated genes associated with Ca 2+ and phosphate homeostasis in rats fed control or phytate-supplemented diets (n = 8 per group). ( B ) Results of immunoblot analyses and protein levels of renal aKlotho, NHERF1, and NaPi-2a after 2 weeks for rats fed control, HP- LCa 2+ , and HP-HCa 2+ diets. ( C ) The levels of renal proteins were quantified using ImageJ software (n = 6 per group). ( D, E ) Time course analysis of serum levels of intact FGF23 ( D ) and C-terminal FGF23 ( E ) in rats fed control, HP-LCa 2+ , and HP-HCa 2+ diets. ( F ) Representative immunohistochemical staining of EDTA-decalcified femur sections for FGF23 from rats fed control, HP-LCa 2+ , and HP-HCa 2+ diets. All data are presented as the mean ± SD of each group (n = 3–8 per group). *, p<0.05; **, p<0.01; ***, p<0.001, compared with controls.

Article Snippet: Rat intact PTH (Immutopics, San Clemente, CA, USA), human intact PTH (Abcam), soluble RANKL (Immundiagnostik, Bensheim, Germany), osteoprotegerin (Alpco Immunoassay, Salem, NH, USA), rat intact and C-terminal FGF23 (Elabscience, Houston, TX, USA), and human intact FGF23 (Elabscience) were measured using enzyme-linked immunosorbent assay.

Techniques: Expressing, Control, Western Blot, Software, Immunohistochemical staining, Staining

Journal: eLife

Article Title: High-phytate/low-calcium diet is a risk factor for crystal nephropathies, renal phosphate wasting, and bone loss

doi: 10.7554/eLife.52709

Figure Lengend Snippet:

Article Snippet: Rat intact PTH (Immutopics, San Clemente, CA, USA), human intact PTH (Abcam), soluble RANKL (Immundiagnostik, Bensheim, Germany), osteoprotegerin (Alpco Immunoassay, Salem, NH, USA), rat intact and C-terminal FGF23 (Elabscience, Houston, TX, USA), and human intact FGF23 (Elabscience) were measured using enzyme-linked immunosorbent assay.

Techniques: Control, Enzyme-linked Immunosorbent Assay, RIA Assay, Staining, Colorimetric Assay, Blocking Assay, Recombinant, In Vitro, Software