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R&D Systems rat anti fgf23
Rat Anti Fgf23, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rat+anti+mouse+fgf23/Mouse+FGF-23+Antibody/pmc12395529-115-6-10
Average 93 stars, based on 45 article reviews

rat anti fgf23 - by Bioz Stars, 2026-09

93/100 stars

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Affinity Purification:

Article Title: Posttranslational processing of FGF23 in osteocytes during the osteoblast to osteocyte transition
Article Snippet: .. Antisera were then applied as follows: affinity-purified rat anti-mouse FGF23 (R&D Systems MAB26291, 1:100); M2 Flag antibody (Sigma F3165, 1:50): sheep anti-rat TGN38 (Serotec AHP499G, 1:100); affinity-purified rabbit polyclonal furin antibody (Affinity Bioreagents PA1-062, 1:100); affinity-purified sheep anti-human/mouse PC2 (R&D Systems AF6018, 1:100); and/or rabbit anti-human PC5/6 serum at 1:40 (R. Day, U. Sherbrooke). .. After washing, the cells were incubated for 1 hour at room temperature with one or more of the following fluorescence-conjugated secondary antibodies at 1:800 to 1:1000: anti-rat IgG-Cy3, anti-rabbit IgG-Cy3, anti-sheep IgG-Cy5, or anti-rat IgG-Cy5 (Jackson Immunoresearch, West Grove, PA) in blocking solution containing DAPI nuclear/DNA stain reagent (1:10,000, ALX-620-050, Axxora LLC, San Diego, CA).

Staining:

Article Title: Calcium and vitamin-D deficiency marginally impairs fracture healing but aggravates posttraumatic bone loss in osteoporotic mice
Article Snippet: The mineral-apposition rate (MAR), bone-formation rate per bone surface (BFR/BS) and mineralized surface per bone surface (MS/BS) were determined in the second lumbar vertebral body (d23; 0.6 × 0.6 mm centered region) using Osteomeasure® image analysis (OsteoMetrics). .. Paraffin-embedded sections of fractured femurs (d23) were immunohistochemically stained for FGF23 and fibroblast growth factor receptor 1 (FGFR1) using the following antibodies and dilutions: rat anti-mouse FGF23, 1:100 (MAB26291, R&D systems Inc., Minneapolis, USA), rabbit anti-mouse FGFR1/CD331, 1:50 (PA5-25979, Invitrogen, Thermo Fisher Scientific, Waltham, USA), goat anti-rabbit immunoglobulin G (IgG) (H + L) biotin-conjugated, 1:100 (B2770, Invitrogen), and goat anti-rat IgG (H + L) biotin-conjugated, 1:100 (A10517, Invitrogen). ..

Image Search Results

Original photographs of the opened stomach of a CD-fed mouse with normal gastric mucosa ( A ) and a mouse after 12 weeks on HFD with mucosal abnormalities and poly-like lesions (dashed circle) ( B ). C Original gel photograph showing the products of gastric Fgf23 -specific transcript amplification in the presence (+RT) or absence (-RT) of reverse transcriptase using pooled RNA samples from each group (CD: 44 per group; HFD: 6 per group). D Arithmetic means ± SEM of relative gastric Fgf23 transcript levels normalized to Rpl8 (CD: n = 5; HFD: n = 6; Mann-Whitney U test). E Original Western blot showing FGF23 abundance in homogenates from the proximal stomach of CD- and HFD-fed mice using bone as positive control (upper panel) and total protein staining as a loading control (lower panel). Size of marker (in kDa) is shown on the right. F Densitometric analysis of FGF23 protein. Data are expressed as fold change of FGF23 protein expression relative to total protein in CD- versus HFD-fed mice and shown as means ± SEM (CD: n = 4; HFD: n = 4; Welch’s t -test). Localization of H + /K + -ATPase (red) and FGF23 (green) in the oxyntic mucosa of the proximal stomach of a CD-fed mouse ( G ) and of a mouse on HFD ( H ). Localization of TFF1 (red) and FGF23 (green) in a CD-fed mouse ( I ) and a mouse on HFD ( J ). A dashed line encircles distended glands, hashmarks point to interglandular spaces, arrowheads to surface epithelium and asterisks to blood vessels (CD: n = 3; HFD: n = 4). G – J Sections were counterstained with DAPI. Scale bars: 100 μm. *p < 0.05. bp base pairs; CD control diet; FGF23 fibroblast growth factor 23; H + /K + -ATPase hydrogen potassium-ATPase; HFD high fat diet; Rpl8 ribosomal protein L8; RT reverse transcriptase; TFF1 trefoil factor 1.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: Original photographs of the opened stomach of a CD-fed mouse with normal gastric mucosa ( A ) and a mouse after 12 weeks on HFD with mucosal abnormalities and poly-like lesions (dashed circle) ( B ). C Original gel photograph showing the products of gastric Fgf23 -specific transcript amplification in the presence (+RT) or absence (-RT) of reverse transcriptase using pooled RNA samples from each group (CD: 44 per group; HFD: 6 per group). D Arithmetic means ± SEM of relative gastric Fgf23 transcript levels normalized to Rpl8 (CD: n = 5; HFD: n = 6; Mann-Whitney U test). E Original Western blot showing FGF23 abundance in homogenates from the proximal stomach of CD- and HFD-fed mice using bone as positive control (upper panel) and total protein staining as a loading control (lower panel). Size of marker (in kDa) is shown on the right. F Densitometric analysis of FGF23 protein. Data are expressed as fold change of FGF23 protein expression relative to total protein in CD- versus HFD-fed mice and shown as means ± SEM (CD: n = 4; HFD: n = 4; Welch’s t -test). Localization of H + /K + -ATPase (red) and FGF23 (green) in the oxyntic mucosa of the proximal stomach of a CD-fed mouse ( G ) and of a mouse on HFD ( H ). Localization of TFF1 (red) and FGF23 (green) in a CD-fed mouse ( I ) and a mouse on HFD ( J ). A dashed line encircles distended glands, hashmarks point to interglandular spaces, arrowheads to surface epithelium and asterisks to blood vessels (CD: n = 3; HFD: n = 4). G – J Sections were counterstained with DAPI. Scale bars: 100 μm. *p < 0.05. bp base pairs; CD control diet; FGF23 fibroblast growth factor 23; H + /K + -ATPase hydrogen potassium-ATPase; HFD high fat diet; Rpl8 ribosomal protein L8; RT reverse transcriptase; TFF1 trefoil factor 1.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat anti-fibroblast growth factor 23 (FGF23) (MAB26291, R&D systems, Minneapolis, MN, USA), mouse anti-hydrogen potassium-ATPase (H + /K + -ATPase) as parietal cell marker (sc-374094, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse anti-platelet endothelial cell adhesion molecule (PECAM/CD31) for endothelial cells (sc-376764, Santa Cruz Biotechnology), rabbit anti-trefoil factor 1 (TFF1) for mucus-producing cells (GTX121461, GeneTex, Irvine, CA, USA), rabbit anti-α-smooth muscle actin (α-SMA) to visualize pericytes and smooth muscle cells (ab5694, Abcam, Cambridge, England), the leukocyte marker rat anti-CD45 to detect immune infiltrates (550539, BD Biosciences, Heidelberg, Germany), and rabbit anti-IL-1β for labeling of infiltrating and epithelial cells (at a dilution of 1:200; P420B, Thermo Fisher Scientific, Darmstadt, Germany).

Techniques: Amplification, Reverse Transcription, MANN-WHITNEY, Western Blot, Positive Control, Staining, Control, Marker, Expressing

Epithelial lesions visualized by TFF1 staining in CD-fed mice ( A ) and mice on HFD ( B ). Coimmunolabeling of FGF23 (green) and TFF1 (red) in patchy disorganized glands in the stomach of a CD-fed mouse ( C ) and of a mouse on HFD ( D ). Insets: PAS staining verifies the presence of mucin in gastric lesions. Extensive purple color indicates pronounced mucus production, most prominently in elongated foveolae and some dilated glands in the midportion of the mucosa. E - G Higher magnification of the stomach from a mouse on HFD. (CD: n = 3; HFD: n = 4). H Arithmetic means ± SEM of FGF23-/TFF1-positive areas expressed as proportion of total mucosal thickness (CD: n = 3; HFD: n = 4; student’s t -test). I Arithmetic means ± SEM of the number of cells positive for the indicated markers counted in mucosal lesions (CD: n = 3; HFD: n = 4; cells positive for FGF23/TFF1 and TFF1: student’s t -test, cells positive for FGF23: Welch’s t -test). A dashed line encircles epithelial lesions. Sections were counterstained with DAPI. Scale bars: 100 µm ( A , B ), 50 μm ( C , D ), 10 μm ( E – G ). *p < 0.05; **p <0.01. CD control diet, FGF23 fibroblast growth factor 23, HFD high fat diet, PAS periodic acid-Schiff, TFF1, trefoil factor 1.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: Epithelial lesions visualized by TFF1 staining in CD-fed mice ( A ) and mice on HFD ( B ). Coimmunolabeling of FGF23 (green) and TFF1 (red) in patchy disorganized glands in the stomach of a CD-fed mouse ( C ) and of a mouse on HFD ( D ). Insets: PAS staining verifies the presence of mucin in gastric lesions. Extensive purple color indicates pronounced mucus production, most prominently in elongated foveolae and some dilated glands in the midportion of the mucosa. E - G Higher magnification of the stomach from a mouse on HFD. (CD: n = 3; HFD: n = 4). H Arithmetic means ± SEM of FGF23-/TFF1-positive areas expressed as proportion of total mucosal thickness (CD: n = 3; HFD: n = 4; student’s t -test). I Arithmetic means ± SEM of the number of cells positive for the indicated markers counted in mucosal lesions (CD: n = 3; HFD: n = 4; cells positive for FGF23/TFF1 and TFF1: student’s t -test, cells positive for FGF23: Welch’s t -test). A dashed line encircles epithelial lesions. Sections were counterstained with DAPI. Scale bars: 100 µm ( A , B ), 50 μm ( C , D ), 10 μm ( E – G ). *p < 0.05; **p <0.01. CD control diet, FGF23 fibroblast growth factor 23, HFD high fat diet, PAS periodic acid-Schiff, TFF1, trefoil factor 1.

Techniques: Staining, Control

Distribution of blood vessels in the proximal glandular stomach from a CD-fed mouse ( A ) and a mouse on HFD ( B ). Staining for FGF23 ( C, F ), PECAM ( D ), and α-SMA ( G ) and the merge of FGF23 and PECAM ( E ) or FGF23 and α-SMA ( H ) staining in the proximal glandular stomach from a mouse on HFD. ( A – H : CD: n = 3; HFD: n = 4). Arithmetic means ± SEM of the density of FGF23-/PECAM-positive blood vessels (CD: n = 3; HFD: n = 4; student’s t -test) ( I ) and of the number of endothelial cells costained for FGF23 and PECAM (CD: n = 3; HFD: n = 4; Welch’s t -test) ( J ). Asterisks point to blood vessels. Sections were counterstained with DAPI. Scale bars: 100 μm ( A , B ), 10 μm ( C – H ). *p < 0.05; ***p < 0.001. α-SMA α-smooth muscle actin, CD control diet, FGF23 fibroblast growth factor 23, HFD high fat diet, PECAM platelet endothelial cell adhesion molecule.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: Distribution of blood vessels in the proximal glandular stomach from a CD-fed mouse ( A ) and a mouse on HFD ( B ). Staining for FGF23 ( C, F ), PECAM ( D ), and α-SMA ( G ) and the merge of FGF23 and PECAM ( E ) or FGF23 and α-SMA ( H ) staining in the proximal glandular stomach from a mouse on HFD. ( A – H : CD: n = 3; HFD: n = 4). Arithmetic means ± SEM of the density of FGF23-/PECAM-positive blood vessels (CD: n = 3; HFD: n = 4; student’s t -test) ( I ) and of the number of endothelial cells costained for FGF23 and PECAM (CD: n = 3; HFD: n = 4; Welch’s t -test) ( J ). Asterisks point to blood vessels. Sections were counterstained with DAPI. Scale bars: 100 μm ( A , B ), 10 μm ( C – H ). *p < 0.05; ***p < 0.001. α-SMA α-smooth muscle actin, CD control diet, FGF23 fibroblast growth factor 23, HFD high fat diet, PECAM platelet endothelial cell adhesion molecule.

Techniques: Staining, Control

Staining for FGF23 in the gastric interglandular spaces of a CD-fed mouse ( A ) and a mouse on HFD ( B ). Immunostaining for FGF23 ( C, F, I ), PECAM ( D ), α-SMA ( G, J ), and merge of FGF23 and PECAM ( E ) or FGF23 and α-SMA ( H, K ) staining in interglandular spaces of the corpus ( C - H ) and around cardia glands ( I – K ) from a mouse on HFD. (CD: n = 3; HFD: n = 4). Asterisks point to blood vessels, white arrowheads to stromal endothelial cells, and yellow arrowheads to myofibroblasts. Sections were counterstained with DAPI. Scale bars: 100 μm ( A , B ), 20 μm ( C – K ). α-SMA α-smooth muscle actin, CD control diet, FGF23 fibroblast growth factor 23, HFD high fat diet, PECAM platelet endothelial cell adhesion molecule.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: Staining for FGF23 in the gastric interglandular spaces of a CD-fed mouse ( A ) and a mouse on HFD ( B ). Immunostaining for FGF23 ( C, F, I ), PECAM ( D ), α-SMA ( G, J ), and merge of FGF23 and PECAM ( E ) or FGF23 and α-SMA ( H, K ) staining in interglandular spaces of the corpus ( C - H ) and around cardia glands ( I – K ) from a mouse on HFD. (CD: n = 3; HFD: n = 4). Asterisks point to blood vessels, white arrowheads to stromal endothelial cells, and yellow arrowheads to myofibroblasts. Sections were counterstained with DAPI. Scale bars: 100 μm ( A , B ), 20 μm ( C – K ). α-SMA α-smooth muscle actin, CD control diet, FGF23 fibroblast growth factor 23, HFD high fat diet, PECAM platelet endothelial cell adhesion molecule.

Techniques: Staining, Immunostaining, Control

A Lipid deposits stained by Nile red in metaplastic glands. The right panels indicate the framed area with droplets in FGF23 + cells at higher magnification. B The upper panel shows FGF23 + surface mucus and desquamated cells, displaying strong Nile red staining (lower panel) in the contiguous sections. (CD: n = 3; HFD: n = 4). Arithmetic means ± SEM of gastric Muc2 ( C ), Il-6 ( D ), Il-11 ( E ), Tnf-α ( F ) and Dmp1 ( G ) transcript abundance relative to Rpl8 in CD- and HFD-fed mice (n = 5-6 per group; C , D , G student’s t -test, E , F Mann-Whitney U test). FGF23 + metaplastic gland area ( H ) surrounded by CD45 + immune cells ( I ) of consecutive sections. Contiguous sections showing dilated FGF23 + interconnected blood vessels ( J ) enwrapped by CD45 + infiltrates ( K ). FGF23 + disorganized glands ( L ) contained IL-1β + inflammatory and epithelial cells ( M ). (CD: n = 3; HFD: n = 4). Sections were counterstained with DAPI. Scale bars ( A , B , H – M ): 50 μm. *p < 0.05. CD control diet, CD45 cluster of differentiation 45, Dmp1 dentin matrix protein 1 , FGF23 fibroblast growth factor 23, HFD high fat diet, IL-1β interleukin 1 β, Il-6 interleukin 6, Il-11 interleukin 11, Muc2 mucin 2, Rpl8 ribosomal protein L8, Tnf-α tumor necrosis factor-α.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: A Lipid deposits stained by Nile red in metaplastic glands. The right panels indicate the framed area with droplets in FGF23 + cells at higher magnification. B The upper panel shows FGF23 + surface mucus and desquamated cells, displaying strong Nile red staining (lower panel) in the contiguous sections. (CD: n = 3; HFD: n = 4). Arithmetic means ± SEM of gastric Muc2 ( C ), Il-6 ( D ), Il-11 ( E ), Tnf-α ( F ) and Dmp1 ( G ) transcript abundance relative to Rpl8 in CD- and HFD-fed mice (n = 5-6 per group; C , D , G student’s t -test, E , F Mann-Whitney U test). FGF23 + metaplastic gland area ( H ) surrounded by CD45 + immune cells ( I ) of consecutive sections. Contiguous sections showing dilated FGF23 + interconnected blood vessels ( J ) enwrapped by CD45 + infiltrates ( K ). FGF23 + disorganized glands ( L ) contained IL-1β + inflammatory and epithelial cells ( M ). (CD: n = 3; HFD: n = 4). Sections were counterstained with DAPI. Scale bars ( A , B , H – M ): 50 μm. *p < 0.05. CD control diet, CD45 cluster of differentiation 45, Dmp1 dentin matrix protein 1 , FGF23 fibroblast growth factor 23, HFD high fat diet, IL-1β interleukin 1 β, Il-6 interleukin 6, Il-11 interleukin 11, Muc2 mucin 2, Rpl8 ribosomal protein L8, Tnf-α tumor necrosis factor-α.

Techniques: Staining, MANN-WHITNEY, Control

Arithmetic means ± SEM of relative gastric transcript levels of Fgf23 ( A ), Sox2 ( B ), Il-6 ( C ), and Il-11 ( D ) normalized to Rpl8 after one week on HFD compared to CD-fed mice (CD: n = 5; HFD: n = 4; student’s t -test). Contiguous sections showing the formation of narrow-lumened FGF23 + interconnected blood vessels ( E ) surrounded by CD45 + cells ( F ). FGF23 + blood vessel ( G ) containing Nile red-stained lipid deposits ( H ) of contiguous sections. (CD: n = 5; HFD: n = 4). Sections were counterstained with DAPI. Scale bars ( E – H ): 50 μm. *p < 0.05; **p < 0.01. bp base pairs, CD control diet, CD45 cluster of differentiation 45, Fgf23 fibroblast growth factor 23, HFD high fat diet, Il-6 interleukin 6, Il-11 interleukin 11, Rpl8 ribosomal protein L8, Sox2 SRY-box transcription factor 2.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: Arithmetic means ± SEM of relative gastric transcript levels of Fgf23 ( A ), Sox2 ( B ), Il-6 ( C ), and Il-11 ( D ) normalized to Rpl8 after one week on HFD compared to CD-fed mice (CD: n = 5; HFD: n = 4; student’s t -test). Contiguous sections showing the formation of narrow-lumened FGF23 + interconnected blood vessels ( E ) surrounded by CD45 + cells ( F ). FGF23 + blood vessel ( G ) containing Nile red-stained lipid deposits ( H ) of contiguous sections. (CD: n = 5; HFD: n = 4). Sections were counterstained with DAPI. Scale bars ( E – H ): 50 μm. *p < 0.05; **p < 0.01. bp base pairs, CD control diet, CD45 cluster of differentiation 45, Fgf23 fibroblast growth factor 23, HFD high fat diet, Il-6 interleukin 6, Il-11 interleukin 11, Rpl8 ribosomal protein L8, Sox2 SRY-box transcription factor 2.

Techniques: Staining, Control

A – C Blood vessels in the stomach of a normal-weight subject showing endothelial cells faintly immunoreactive for FGF23 ( A ) but strongly for PECAM ( B ). D Gastric blood vessels with interconnections at the base of gastric glands in patients with obesity. E – G Gastric endothelial cells of a patient with obesity display strong immunoreactivity for FGF23 ( E ) and PECAM ( F ). (Normal-weight: n = 6; patients with obesity: n = 6). A dotted line marks interconnected blood vessels. Sections were counterstained with DAPI. Scale bars: 20 μm ( A - C , E - G ), 50 μm ( D ). FGF23 fibroblast growth factor 23, PECAM platelet endothelial cell adhesion molecule.

Journal: International Journal of Obesity (2005)

Article Title: High fat diet induces gastric production of fibroblast growth factor 23 (FGF23)

doi: 10.1038/s41366-025-01808-3

Figure Lengend Snippet: A – C Blood vessels in the stomach of a normal-weight subject showing endothelial cells faintly immunoreactive for FGF23 ( A ) but strongly for PECAM ( B ). D Gastric blood vessels with interconnections at the base of gastric glands in patients with obesity. E – G Gastric endothelial cells of a patient with obesity display strong immunoreactivity for FGF23 ( E ) and PECAM ( F ). (Normal-weight: n = 6; patients with obesity: n = 6). A dotted line marks interconnected blood vessels. Sections were counterstained with DAPI. Scale bars: 20 μm ( A - C , E - G ), 50 μm ( D ). FGF23 fibroblast growth factor 23, PECAM platelet endothelial cell adhesion molecule.

Techniques:

Figure 5. Immunolocalization of FGF23, MEPE, and osteopontin in the control and PTH-administered metaphyseal trabeculae. Panels (A–C,D–F,G–I) are immunodetection of FGF23, MEPE, and osteo- pontin, respectively. Panels (A,D,G) show the control metaphyses, while panels (B,E,H) and (C,F,I) are from PTH-administered trabeculae with regimens of 2 times or 4 times/day, respectively. FGF23 is observed mainly in osteocytes of control specimens (A), while FGF23 can be seen not only in osteocytes but also in osteoblasts after the PTH administration (arrows in B,C). MEPE is observable in osteocytes after the PTH treatment (E,F), while it is hardly detected in the control specimens (D). Note intense osteopontin-immunoreactive cement lines (arrows in H,I) in the PTH-administered specimens compared to the control counterparts (G). tb: trabecula. Bars (A–I): 30 µm.

Journal: Medicina (Kaunas, Lithuania)

Article Title: Immunolocalization of Enzymes/Membrane Transporters Related to Bone Mineralization in the Metaphyses of the Long Bones of Parathyroid-Hormone-Administered Mice.

doi: 10.3390/medicina59061179

Figure Lengend Snippet: Figure 5. Immunolocalization of FGF23, MEPE, and osteopontin in the control and PTH-administered metaphyseal trabeculae. Panels (A–C,D–F,G–I) are immunodetection of FGF23, MEPE, and osteo- pontin, respectively. Panels (A,D,G) show the control metaphyses, while panels (B,E,H) and (C,F,I) are from PTH-administered trabeculae with regimens of 2 times or 4 times/day, respectively. FGF23 is observed mainly in osteocytes of control specimens (A), while FGF23 can be seen not only in osteocytes but also in osteoblasts after the PTH administration (arrows in B,C). MEPE is observable in osteocytes after the PTH treatment (E,F), while it is hardly detected in the control specimens (D). Note intense osteopontin-immunoreactive cement lines (arrows in H,I) in the PTH-administered specimens compared to the control counterparts (G). tb: trabecula. Bars (A–I): 30 µm.

Article Snippet: For FGF23, after pre-incubation with 1% BSA-PBS for 30 min at RT, sections were incubated with rat anti-FGF23 antibodies (R&D Systems Inc., Minneapolis, MN, USA) at a dilution of 1:100 for 2 h [37] and then incubated with HRP-conjugated anti-rat IgG (Chemicon International Inc., Temecula, CA, USA) at a dilution of 1:100 for 1 h. For ENPP1, sections were incubated with goat polyclonal anti-ENPP1 (Everest Biotech Ltd., Oxfordshire, UK) at a dilution of 1:200 for 1 h and then Medicina 2023, 59, 1179 4 of 16 incubated with HRP-conjugated rabbit anti-goat IgG (American Qualex Scientific Products, Inc., San Clemente, CA, USA).

Techniques: Control, Immunodetection

Figure 6. The gene expression proﬁles of Tnalp, Enpp1, Phospho1, Ank, Mepe, Phex, Fgf23, Fgfr1, and klotho assessed by RT-PCR in the control and PTH-administered bones. Panel (A) reveals RT-PCR results indicating the gene expressions of Tnalp, Enpp1, Phospho1, and Ank, while panel (B) displays the expressions of Mepe, Phex, Fgf23, Fgfr1, and klotho examined by RT-PCR.

Journal: Medicina (Kaunas, Lithuania)

Article Title: Immunolocalization of Enzymes/Membrane Transporters Related to Bone Mineralization in the Metaphyses of the Long Bones of Parathyroid-Hormone-Administered Mice.

doi: 10.3390/medicina59061179

Figure Lengend Snippet: Figure 6. The gene expression proﬁles of Tnalp, Enpp1, Phospho1, Ank, Mepe, Phex, Fgf23, Fgfr1, and klotho assessed by RT-PCR in the control and PTH-administered bones. Panel (A) reveals RT-PCR results indicating the gene expressions of Tnalp, Enpp1, Phospho1, and Ank, while panel (B) displays the expressions of Mepe, Phex, Fgf23, Fgfr1, and klotho examined by RT-PCR.

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Control

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.

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.

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.

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.

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.

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.

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.

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.

Techniques: Gene Expression, Activation Assay

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