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: Fig. 1 HFD feeding induces gastric lesions, Fgf23 upregulation and FGF23+ cell expansion in the proximal glandular stomach. 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 antifibroblast 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: Reverse Transcription, MANN-WHITNEY, Western Blot, Positive Control, Staining, Control, Marker, Expressing

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: Fig. 2 HFD-induced appearance of FGF23-expressing mucus-like cells in gastric lesions. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Expressing, Staining, Control

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: Fig. 3 HFD-induced blood vessel formation associated with numerous FGF23-expressing endothelial cells. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Expressing, Staining, Control

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: Fig. 4 HFD-feeding enhances FGF23 immunoreactivity within periglandular cells of the gastric stroma. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Staining, Immunostaining, Control

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: Fig. 5 Signs of lipotoxicity, impaired mucosal integrity and local inflammation in FGF23+ mucosal areas. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Staining, MANN-WHITNEY, Control

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: Fig. 6 One week on HFD induces upregulation of FGF23 and mild gastric lesions harboring FGF23+ cells, CD45+ immune cells and lipid accumulation. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Staining, Control

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: Fig. 7 Endothelial cells in the human stomach express FGF23. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Peritoneal repairing cells: a type of bone marrow derived progenitor cells involved in mesothelial regeneration

doi: 10.1111/j.1582-4934.2010.01087.x

Figure Lengend Snippet: Antigen immunolocalization in adherent cells obtained from peritoneal lavages 48 hrs after injury of the peritoneal wall. (A–C) show cells obtained from normal BALB/c mice whereas (D)–(F) show cells from mice reconstituted with GFP-expressing bone marrow. (A) Virtually all the CD45 + cells show a perinuclear dot-like pattern of cytokeratin immunoreactivity. Note a strongly CD45 – /cytokeratin + immunoreactive cell, probably a delaminated mesothelial cell (arrow). (B) The perinuclear dot-like pattern of cytokeratin immunoreactivity is also present in mesothelial cells migrating from omentum explants, which were used as positive controls (arrows). Other cells still show an extended cytokeratin cytoskeleton (arrowhead). (C) Mesothelin immunoreactivity colocalized with CD68. Note the cytoplasmic and perinuclear CD68 localization. (D) Triple localization of mesothelin, cytokeratin and GFP. This immunostaining revealed different phenotypes, including triple positive cells (arrowheads), mesothelin + /cytokeratin + , GFP – cells which probably are delaminated mesothelial cells (white arrows) and GFP + , mesothelin – /cytokeratin – cells (yellow arrow). (E) The fibroblast marker FSP1 was expressed in many GFP + cells (white arrows), as well as in GFP – cells (yellow arrows). Other GFP + cells were negative for FSP1 (arrowhead). (F) Negative control incubated with isotype primary antibodies and with the same secondary antibodies as used in the rest of the figures.

Article Snippet: Primary antibodies used were: polyclonal rabbit anti-cytokeratin (DAKO, Z0622) diluted 1:200; polyclonal rabbit anti-fibroblast specific protein (FSP)1 (A kind gift from Dr. Eric Neilson, Vanderbilt University School of Medicine) diluted 1:100; rat antimouse CD68 (AbD Serotec, Dusseldorf, Germany, MCA1957BT, Clone FA-11) diluted 1:100; polyclonal rabbit anti-mesothelin (A kind gift from Dr. Ira Pastan, NCI-NIH) diluted 1:200; mouse anti-SMC α-actin (Sigma A2547, Clone 1A4) diluted 1:200; goat polyclonal anti-procollagen I (Santa Cruz, Biotechnologies, Heidelberg, Germany; SC-8787) diluted 1:100.

Techniques: Expressing, Immunostaining, Marker, Negative Control, Incubation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Peritoneal repairing cells: a type of bone marrow derived progenitor cells involved in mesothelial regeneration

doi: 10.1111/j.1582-4934.2010.01087.x

Figure Lengend Snippet: Expression of markers on macrophages, PRC, fibrocytes and fibroblasts according our own results and data from [ 23 ]

Article Snippet: Primary antibodies used were: polyclonal rabbit anti-cytokeratin (DAKO, Z0622) diluted 1:200; polyclonal rabbit anti-fibroblast specific protein (FSP)1 (A kind gift from Dr. Eric Neilson, Vanderbilt University School of Medicine) diluted 1:100; rat antimouse CD68 (AbD Serotec, Dusseldorf, Germany, MCA1957BT, Clone FA-11) diluted 1:100; polyclonal rabbit anti-mesothelin (A kind gift from Dr. Ira Pastan, NCI-NIH) diluted 1:200; mouse anti-SMC α-actin (Sigma A2547, Clone 1A4) diluted 1:200; goat polyclonal anti-procollagen I (Santa Cruz, Biotechnologies, Heidelberg, Germany; SC-8787) diluted 1:100.

Techniques: Expressing

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: Fig. 1 HFD feeding induces gastric lesions, Fgf23 upregulation and FGF23+ cell expansion in the proximal glandular stomach. 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 antifibroblast 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: Reverse Transcription, MANN-WHITNEY, Western Blot, Positive Control, Staining, Control, Marker, Expressing

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: Fig. 2 HFD-induced appearance of FGF23-expressing mucus-like cells in gastric lesions. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Expressing, Staining, Control

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: Fig. 5 Signs of lipotoxicity, impaired mucosal integrity and local inflammation in FGF23+ mucosal areas. 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.

Article Snippet: For murine sections, the following antibodies at a dilution of 1:400 (except for IL-1β) were used: rat antifibroblast 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: Staining, MANN-WHITNEY, Control

Journal: Particle and Fibre Toxicology

Article Title: Stat-6 signaling pathway and not Interleukin-1 mediates multi-walled carbon nanotube-induced lung fibrosis in mice: insights from an adverse outcome pathway framework

doi: 10.1186/s12989-017-0218-0

Figure Lengend Snippet: Fibrotic disease is not attenuated 28 days after MWCNT exposure. C57BL/6 and IL-1R1 KO mice were intratracheally administered 162μg of MWCNTs, and samples were collected 24 h and 28 days later. a The pro-fibrotic genes CCL2, osteopontin (OPN), and total TGF-β levels including the active form were measured by ELISA. b Representative histology images of the diseased area of lungs 28 days after MWCNT instillation compare WT and IL-1R1 KO mice. These images were obtained from slides stained with H&E, Masson Trichrome for collagen deposition (blue), and immune-staining for Vimentin, a surface marker of fibroblasts (brown). The scale bar represents 50 μm. c Entire longitudinal cross sections of the lungs were imaged and the disease area verses the total lung area were determined to quantify the pathology in H&E stained samples 28 days post exposure. d Representative images of pathology were taken of the Masson trichrome stained slides and the amount of collagen positive stain was quantified and normalized as a percent of area imaged. Non-diseased areas from exposed mice were used as controls. e Soluble collagen was measured in the BAL fluid at the 28 day time point, and f total protein was measured in BAL as well. Data represent mean ± SEM. n = 4–5. Statistical analysis was performed using two-way ANOVA. * p < 0.05, NS = not significant, †statistical interaction with p < 0.05, and NI = no statistical interaction

Article Snippet: Additionally, immunohistochemistry for the fibroblast marker vimentin (Rabbit vimentin antibody, Catalogue #5741, Clone D21H3, 1:100 dilutions; Cell Signaling Technologies, Danvers, MA) was also performed on formalin-fixed, paraffin embedded lung tissue sections.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Marker

Journal: Particle and Fibre Toxicology

Article Title: Stat-6 signaling pathway and not Interleukin-1 mediates multi-walled carbon nanotube-induced lung fibrosis in mice: insights from an adverse outcome pathway framework

doi: 10.1186/s12989-017-0218-0

Figure Lengend Snippet: Fibrotic disease but not traditional fibrotic markers are attenuated 28 days after MWCNT exposure. C57BL/6 and STAT6 KO mice were intratracheally administered 162 μg of MWCNTs, and samples were collected 24 h and 28 days later. a The pro-fibrotic genes CCL2, osteopontin (OPN), and total TGF-β levels including the active form were measured by ELISA. b Representative histology images of the diseased area of lungs 28 days after MWCNT instillation compare WT and STAT6 KO mice. These images were obtained from slides stained with H&E, Masson Trichrome for collagen deposition (blue), and immune-staining for Vimentin, a surface marker of fibroblasts (brown). The scale bar represents 50 μm. c Entire longitudinal cross sections of the lungs were imaged and the disease area versus the total lung area was determined to quantify the pathology in H&E stained samples 28 days post exposure. d Representative images of pathology were taken of the Masson trichrome stained slides, and the amount of collagen positive stain was quantified and normalized as a percent of area imaged. Non-diseased areas from exposed mice were imaged as controls. e Soluble collagen was measured in the BAL fluid at the 28 day time point, and f total protein was measured in BAL as well. Data represent mean ± SEM. n = 4–5. Statistical analysis was performed using two-way ANOVA. *p < 0.05, NS = not significant, †statistical interaction with p < 0.05, and NI = no statistical interaction

Article Snippet: Additionally, immunohistochemistry for the fibroblast marker vimentin (Rabbit vimentin antibody, Catalogue #5741, Clone D21H3, 1:100 dilutions; Cell Signaling Technologies, Danvers, MA) was also performed on formalin-fixed, paraffin embedded lung tissue sections.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Marker