Journal: Molecular Medicine Reports

Article Title: Sonic hedgehog protein regulates fibroblast growth factor 8 expression in metanephric explant culture from BALB/c mice: Possible mechanisms associated with renal morphogenesis

doi: 10.3892/mmr.2016.5614

Figure Lengend Snippet: Primer sequences used for polymerase chain reaction.

Article Snippet: Subsequently, sections were incubated overnight at 4°C with polyclonal rabbit anti-Fgf8 (dilution, 1:100; cat. no. ABIN1107218; Bioss Inc., Woburn, MA, USA) and polyclonal rabbit anti-Fgf10 (dilution, 1:100; cat. no. ABIN392510; Bioss Inc.) primary antibodies.

Techniques:

Journal: Molecular Medicine Reports

Article Title: Sonic hedgehog protein regulates fibroblast growth factor 8 expression in metanephric explant culture from BALB/c mice: Possible mechanisms associated with renal morphogenesis

doi: 10.3892/mmr.2016.5614

Figure Lengend Snippet: mRNA expression levels of (A) SHH, (B) Fgf8 and (C) Fgf10 in normal BALB/c mouse embryonic kidney tissue explants at E11.5, E12.5, E13.5 and E14.5 gestational ages. For each sample, the mRNA expression levels were normalized to GAPDH mRNA expression (n=7 for each treatment group ). * P<0.05, **P<0.01 and *** P<0.001 vs. E11.5 group. SHH, sonic hedgehog; Fgf, fibroblast growth factor; E, embryonic day.

Article Snippet: Subsequently, sections were incubated overnight at 4°C with polyclonal rabbit anti-Fgf8 (dilution, 1:100; cat. no. ABIN1107218; Bioss Inc., Woburn, MA, USA) and polyclonal rabbit anti-Fgf10 (dilution, 1:100; cat. no. ABIN392510; Bioss Inc.) primary antibodies.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Sonic hedgehog protein regulates fibroblast growth factor 8 expression in metanephric explant culture from BALB/c mice: Possible mechanisms associated with renal morphogenesis

doi: 10.3892/mmr.2016.5614

Figure Lengend Snippet: mRNA expression levels of (A) Fgf8 and (B) Fgf10 mRNA levels in BALB/c mouse kidney tissue explant cultures following treatment with 1% BSA, 1% BSA + SHH and 1% BSA + SHH + cyclopamine for 4 days. Fgf8 and Fgf10 expression levels were normalized to GAPDH mRNA expression levels (n=7 for each treatment group). ** P<0.01 vs. 1% BSA-alone group; ## P<0.01 vs. 1% BSA+SHH group. Fgf, fibroblast growth factor; BSA, bovine serum albumin; SHH, sonic hedgehog.

Article Snippet: Subsequently, sections were incubated overnight at 4°C with polyclonal rabbit anti-Fgf8 (dilution, 1:100; cat. no. ABIN1107218; Bioss Inc., Woburn, MA, USA) and polyclonal rabbit anti-Fgf10 (dilution, 1:100; cat. no. ABIN392510; Bioss Inc.) primary antibodies.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Sonic hedgehog protein regulates fibroblast growth factor 8 expression in metanephric explant culture from BALB/c mice: Possible mechanisms associated with renal morphogenesis

doi: 10.3892/mmr.2016.5614

Figure Lengend Snippet: Analysis of Fgf8 and Fgf10 expression in BALB/c mouse kidney tissue explant cultures following treatment with 1% BSA (control) or 1% BSA + SHH for 4 days. (A) Immunohistochemical analysis and (B) integral optical densities of Fgf8 and Fgf10 expression levels in the different treatment groups. (C) Western blot analysis and (D) quantification of western blot results, showing the protein expression levels of Fgf8 and Fgf10 expression levels in the various groups. The protein expression levels were normalized to β-actin (n=7 for each treatment group). Scale bar=125 µ m. * P<0.05 and ** P<0.01 vs. 1% BSA-alone group; # P<0.05 and ## P<0.01 vs. 1% BSA + SHH group. Fgf, fibroblast growth factor; BSA, bovine serum albumin; SHH, sonic hedgehog.

Article Snippet: Subsequently, sections were incubated overnight at 4°C with polyclonal rabbit anti-Fgf8 (dilution, 1:100; cat. no. ABIN1107218; Bioss Inc., Woburn, MA, USA) and polyclonal rabbit anti-Fgf10 (dilution, 1:100; cat. no. ABIN392510; Bioss Inc.) primary antibodies.

Techniques: Expressing, Immunohistochemical staining, Western Blot

Journal: The FASEB Journal

Article Title: Loss of the RNA Binding Protein HuR in Early Murine Limb Mesenchyme Does Not Affect Development but Leads to Impaired Bone Homeostasis in Adulthood

doi: 10.1096/fj.202500780RR

Figure Lengend Snippet: Effect of germline Cre recombination in HuR fl/fl mice (Elavl1 KO) through maternal inheritance of Prx1‐Cre. (A) Whole‐mount embryos at E13.5 stained with alcian blue. (B) Toluidine blue‐stained sections of E13.5 embryo forelimbs and hindlimbs. (C) Immunofluorescence detection of HuR in limb sections from E13.5 embryos. (D) Brightfield micrographs demonstrating morphology changes in Control and Elavl1 KO embryos isolated from E9.5 to E11.5. Scale bar = 500 μm. (E) Whole‐mount in situ hybridization chain reaction detection of limb bud markers mRNA in E10.5 and E11.5 embryos. Asterisk: Limb bud, Triangle: Apical ectodermal ridge, Arrowhead: The zone of polarizing activity. Scale bar = 150 μm. (F) Whole‐mount immunofluorescence detection of Fgf8 protein in E10.5 embryos. Asterisk: Limb bud, Triangle: Apical ectodermal ridge. Scale bar = 150 μm.

Article Snippet: Embryos were fixed in 4% formaldehyde for 1 h at 4°C, washed, and stored in 0.0025% Triton X‐100 in PBS (PBS‐Tr) at 4° C. Prior to immunofluorescence staining, embryos were permeabilized with 0.25% Triton X‐100 in PBS for 30 min and then blocked with 2% bovine serum albumin in PBS solution for 1 h before incubation with 1:100 Fgf8 antibody (MAB323, R&D Systems, UK) overnight at 4°C.

Techniques: Staining, Immunofluorescence, Control, Isolation, In Situ Hybridization, Activity Assay

Journal: Development (Cambridge, England)

Article Title: FGF8 is required for cell survival at distinct stages of nephrogenesis and for regulation of gene expression in nascent nephrons.

doi: 10.1242/dev.01944

Figure Lengend Snippet: Fig. 2. Nephrogenesis arrests and cells in the peripheral zone die in the absence of FGF8. Marker analysis in (A,B,Q-T) cryosections or (C-P,U,V) vibratome sections of kidneys at the stages indicated. (A-L,U,V) In situ hybridization for the genes indicated. The Fgf8 (FL) probe we used contained the full-length coding sequence, and therefore detected Fgf8 RNA produced by the Fgf8null allele. (M-P) Immunofluorescence assays for PAX2 (green) to identify the developing nephrons and collecting ducts, co-stained with LysoTracker (LysoT, red) to identify regions containing dying cells. (Q-T) Immunofluorescence assays for PAX2 (green) and Calbindin (CB, blue), which identifies collecting ducts, and for TUNEL staining (red), which detects dying cells. Arrowheads point to nascent nephrons, which are present in normal kidneys and also in Fgf8-MM-KO kidneys at E13.5 (A,B) and E14.5 (Q,R), but not at E16.5 (S,T). Note that the nascent nephrons in Fgf8-MM-KO kidneys (B,R) have formed an epithelial structure surrounding a lumen, i.e. they have reached the renal vesicle stage.

Article Snippet: For experiments on the effects of anti-FGF8 antibody, either normal goat serum (control) or blocking antibody against FGF8 (10 μg/ml, R&D Systems, AF423NA) was added to the culture medium.

Techniques: Marker, In Situ Hybridization, Sequencing, Produced, Staining, TUNEL Assay

Journal: Development (Cambridge, England)

Article Title: FGF8 is required for cell survival at distinct stages of nephrogenesis and for regulation of gene expression in nascent nephrons.

doi: 10.1242/dev.01944

Figure Lengend Snippet: Fig. 3. Signals from the dorsal spinal cord are not sufficient to induce nephrogenesis in Fgf8-deficient metanephric mesenchyme. (A,B) Metanephric mesenchyme (MM) was isolated from E11.5 (A) Fgf8flox/null (control) or (B) Pax3-cre;Fgf8flox/null (Fgf8-MM-KO) littermates and cultured in the presence of dorsal spinal cord. After 48 hours of culture, the samples were processed for immunohistochemistry to detect phospho-Histone H3, which marks cells in mitosis (blue) and E-Cadherin, which marks epithelia (red). The arrowhead in A indicates a region where the tubular nature of the E-CAD-positive structures is particularly evident.

Article Snippet: For experiments on the effects of anti-FGF8 antibody, either normal goat serum (control) or blocking antibody against FGF8 (10 μg/ml, R&D Systems, AF423NA) was added to the culture medium.

Techniques: Isolation, Control, Cell Culture, Immunohistochemistry

Journal: Development (Cambridge, England)

Article Title: FGF8 is required for cell survival at distinct stages of nephrogenesis and for regulation of gene expression in nascent nephrons.

doi: 10.1242/dev.01944

Figure Lengend Snippet: Fig. 4. Kidneys of Fgf8 hypomorphs are smaller than normal, but contain nephrons. (A) Comparison of normal, Fgf8neo/neo (mild hypomorph) and Fgf8neo/null (severe hypomorph) kidneys at E18.5. (B,C) In situ hybridization assay for Wnt4 RNA in vibratome sections of E15.5 kidneys. (D) Immunofluorescence assays at E15.5 for PECAM-positive (red) vasculature and WT1-positive (green) podocyte progenitors in vibratome sections. (D′) A higher magnification view of a renal corpuscle in D.

Article Snippet: For experiments on the effects of anti-FGF8 antibody, either normal goat serum (control) or blocking antibody against FGF8 (10 μg/ml, R&D Systems, AF423NA) was added to the culture medium.

Techniques: Comparison, In Situ Hybridization

Journal: Development (Cambridge, England)

Article Title: FGF8 is required for cell survival at distinct stages of nephrogenesis and for regulation of gene expression in nascent nephrons.

doi: 10.1242/dev.01944

Figure Lengend Snippet: Fig. 5. Nephrons in Fgf8 hypomorphs lack the loop of Henle. (A-L) In situ hybridization assays in vibratome sections of E18.5 normal and Fgf8 hypomorph kidneys, for expression of the genes indicated, identifies podocytes (A-C), proximal tubules (D-F) loop of Henle (G-I) and distal tubules (J-L).

Article Snippet: For experiments on the effects of anti-FGF8 antibody, either normal goat serum (control) or blocking antibody against FGF8 (10 μg/ml, R&D Systems, AF423NA) was added to the culture medium.

Techniques: In Situ Hybridization, Expressing

Journal: Development (Cambridge, England)

Article Title: FGF8 is required for cell survival at distinct stages of nephrogenesis and for regulation of gene expression in nascent nephrons.

doi: 10.1242/dev.01944

Figure Lengend Snippet: Fig. 7. Cells within S-shaped bodies die in Fgf8 hypomorphs. (A-D) Immunofluorescence assay for PAX2 (green), to identify the developing nephrons and collecting ducts, and LysoTracker (LysoT, red) staining to identify regions containing dying cells in vibratome sections of normal and Fgf8 severe hypomorph kidneys at the stages indicated. (A′-D′) Higher magnification views. Arrows indicate regions where Bowman’s capsule progenitors are dying in normal and mutant kidneys; white arrowhead indicates regions where tubule progenitors are dying in the mutant kidney; open arrowheads point to regions in the peripheral zone of the mutant kidney where cells are dying.

Article Snippet: For experiments on the effects of anti-FGF8 antibody, either normal goat serum (control) or blocking antibody against FGF8 (10 μg/ml, R&D Systems, AF423NA) was added to the culture medium.

Techniques: Staining, Mutagenesis

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Evaluation and Immunolocalization of BMP4 and FGF8 in Odontogenic Cyst and Tumors

doi: 10.1155/2018/1204549

Figure Lengend Snippet: Evaluation and immunohistochemical score of stages of odontogenesis.

Article Snippet: The antibody clone used for detection of FGF8 was PA1216 (Boster, Pleasanton, CA, polyclonal antibody).

Techniques: Immunohistochemical staining

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Evaluation and Immunolocalization of BMP4 and FGF8 in Odontogenic Cyst and Tumors

doi: 10.1155/2018/1204549

Figure Lengend Snippet: Evaluation and immunohistochemical score of odontogenic tumors and odontogenic cyst.

Article Snippet: The antibody clone used for detection of FGF8 was PA1216 (Boster, Pleasanton, CA, polyclonal antibody).

Techniques: Immunohistochemical staining

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Evaluation and Immunolocalization of BMP4 and FGF8 in Odontogenic Cyst and Tumors

doi: 10.1155/2018/1204549

Figure Lengend Snippet: Intergroup comparison of expression of BMP4 and FGF8 in odontogenic tumors and odontogenic cyst.

Article Snippet: The antibody clone used for detection of FGF8 was PA1216 (Boster, Pleasanton, CA, polyclonal antibody).

Techniques: Comparison, Expressing

Journal: Oncology letters

Article Title: Systematic analysis of expression profiles and prognostic significance of the FGF gene family in pancreatic adenocarcinoma.

doi: 10.3892/ol.2022.13555

Figure Lengend Snippet: Figure 4. Expression of FGFs in PAAD (Gene Expression Profiling Interactive Analysis). Expression of (A) FGF2, (B) FGF8, (C) FGF9, (D) FGF13, (E) FGF17 and (F) FGF22 in PAAD. *P<0.05. FGF, fibroblast growth factor; PAAD, pancreatic adenocarcinoma; N, normal tissues; T, tumor tissues.

Article Snippet: A total of 50 μl of 5‐10% normal goat serum was added per chip for blocking (1:19 fold dilution) at room temperature for 30 min. Immunohistochemical staining of the paraffin‐embedded tissues was performed using FGF2 (1:200; sc‐74412; Santa Cruz Biotechnology, Inc.) and FGF8 (1:200; 20711‐1‐AP; ProteinTech Group, Inc.) primary anti‐ bodies, anti‐mouse secondary antibodies (1:200; GB23301; Wuhan Servicebio Technology Co., Ltd.), anti‐rabbit secondary antibodies (1:200; GB23303; Wuhan Servicebio Technology Co., Ltd.), and an ABC Elite immunoperoxidase kit (Wuhan Servicebio Technology Co., Ltd.) according to the manufacturer's instructions.

Techniques: Expressing, Gene Expression

Journal: Oncology letters

Article Title: Systematic analysis of expression profiles and prognostic significance of the FGF gene family in pancreatic adenocarcinoma.

doi: 10.3892/ol.2022.13555

Figure Lengend Snippet: Figure 5. Prognostic value of FGFs in PAAD (Gene Expression Profiling Interactive Analysis). Prognostic value of the mRNA expression of (A) FGF2, (B) FGF8, (C) FGF9, (D) FGF13, (E) FGF17 and (F) FGF22 in PAAD. FGF, fibroblast growth factor; PAAD, pancreatic adenocarcinoma; TPM, transcripts per million.

Article Snippet: A total of 50 μl of 5‐10% normal goat serum was added per chip for blocking (1:19 fold dilution) at room temperature for 30 min. Immunohistochemical staining of the paraffin‐embedded tissues was performed using FGF2 (1:200; sc‐74412; Santa Cruz Biotechnology, Inc.) and FGF8 (1:200; 20711‐1‐AP; ProteinTech Group, Inc.) primary anti‐ bodies, anti‐mouse secondary antibodies (1:200; GB23301; Wuhan Servicebio Technology Co., Ltd.), anti‐rabbit secondary antibodies (1:200; GB23303; Wuhan Servicebio Technology Co., Ltd.), and an ABC Elite immunoperoxidase kit (Wuhan Servicebio Technology Co., Ltd.) according to the manufacturer's instructions.

Techniques: Gene Expression, Expressing

Journal: Oncology letters

Article Title: Systematic analysis of expression profiles and prognostic significance of the FGF gene family in pancreatic adenocarcinoma.

doi: 10.3892/ol.2022.13555

Figure Lengend Snippet: Figure 6. Correlations between differentially expressed FGFs and immune cell infiltration (Tumor Immune Estimation Resource). Correlations between the abundance of immune cells and the expression of (A) FGF2, (B) FGF8, (C) FGF9, (D) FGF13, (E) FGF17 and (F) FGF22. FGF, fibroblast growth factor; PAAD, pancreatic adenocarcinoma; TPM, transcripts per million.

Article Snippet: A total of 50 μl of 5‐10% normal goat serum was added per chip for blocking (1:19 fold dilution) at room temperature for 30 min. Immunohistochemical staining of the paraffin‐embedded tissues was performed using FGF2 (1:200; sc‐74412; Santa Cruz Biotechnology, Inc.) and FGF8 (1:200; 20711‐1‐AP; ProteinTech Group, Inc.) primary anti‐ bodies, anti‐mouse secondary antibodies (1:200; GB23301; Wuhan Servicebio Technology Co., Ltd.), anti‐rabbit secondary antibodies (1:200; GB23303; Wuhan Servicebio Technology Co., Ltd.), and an ABC Elite immunoperoxidase kit (Wuhan Servicebio Technology Co., Ltd.) according to the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology letters

Article Title: Systematic analysis of expression profiles and prognostic significance of the FGF gene family in pancreatic adenocarcinoma.

doi: 10.3892/ol.2022.13555

Figure Lengend Snippet: Figure 8. Expression and influence of FGF2 and FGF8 in pancreatic adenocarcinoma. (A) Expression of FGF2 and FGF8 in pancreatic adenocarcinoma. Scale bar, 20 µm. Quantification analysis of IHC for (B) FGF2 (P=0.0313; Wilcoxon's signed rank test) and (C) FGF8 (P=0.0391; Wilcoxon's signed rank test) (*P<0.05). (D) Colony formation assay using MIA Paca‑2 cells treated with DMSO or 10 µM alofanib for 7 days. (E) Colony numbers are presented as the mean ± SD (n=3). *P<0.05. Unpaired Student's t‑test. FGF, fibroblast growth factor; IHC, immunohistochemistry; N, normal tissue; T, tumor tissue.

Article Snippet: A total of 50 μl of 5‐10% normal goat serum was added per chip for blocking (1:19 fold dilution) at room temperature for 30 min. Immunohistochemical staining of the paraffin‐embedded tissues was performed using FGF2 (1:200; sc‐74412; Santa Cruz Biotechnology, Inc.) and FGF8 (1:200; 20711‐1‐AP; ProteinTech Group, Inc.) primary anti‐ bodies, anti‐mouse secondary antibodies (1:200; GB23301; Wuhan Servicebio Technology Co., Ltd.), anti‐rabbit secondary antibodies (1:200; GB23303; Wuhan Servicebio Technology Co., Ltd.), and an ABC Elite immunoperoxidase kit (Wuhan Servicebio Technology Co., Ltd.) according to the manufacturer's instructions.

Techniques: Expressing, Colony Assay, Immunohistochemistry

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Association of circulating PLA2G7 levels with cancer cachexia and assessment of darapladib as a therapy

doi: 10.1002/jcsm.12758

Figure Lengend Snippet: Darapladib treatment was not sufficient to counteract CCx in C26 tumour‐bearing mice despite a strong inhibition of PLA2G7 activity. (A–L) Mice were injected subcutaneously either with PBS (control mice) or C26 cancer cells and treated once daily either with vehicle (PBS mice, white bars, n = 6 animals; C26‐vehicle tumour‐bearing mice, dark grey bars/lines, n = 11 animals) or 50 mg/kg darapladib (C26‐darapladib tumour‐bearing mice, light grey bars/lines, n = 9 animals). (A) Tumour weights. (B) PAF‐AH activity in plasma, (C) tumours and GC muscles. (D) Kaplan–Meier curve depicting the percentage of mice developing cachexia over time. (E) Kinetic of body weight loss during days prior sacrifice. (F) Loss of body weight, and lean and fat mass (expressed as percentage of initial mass). (G) GC muscles, TA muscles, and heart weights. (H) mRNA levels of atrophy and autophagy markers in GC muscle and (I) heart. (J) Epididymal (eWAT), inguinal (iWAT), and brown (BAT) adipose tissues weights. (K) Spleen and lymph nodes weights. (L) Plasma interleukin‐6 (IL‐6) and platelet‐activating factor (PAF) levels ( n = 5 PBS animals, n = 11 C26‐shCTR animals, and n = 9 C26‐sh Pla2g7 animals). Data are mean ± standard error of the mean. Statistical analyses were performed using unpaired t ‐test ( A, C ), unpaired one‐way ANOVA or Kruskal Wallis with Bonferroni or Dunn's post hoc tests, respectively ( B, C, F–L ), paired two‐way ANOVA ( E ), and log‐rank (Mantel–Cox) test ( D ). Tests were two sided. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: CM were diluted with fresh media (1:1, v/v for adipocytes; 3:1, v/v for myotubes) before treatment for 48 h. For darapladib treatment, dimethylsulfoxid (DMSO, vehicle) and 1 μM darapladib (Biorbyt #ORB181231) resuspended in DMSO were freshly added to control or C26 CM before treatment of adipocytes and myotubes.

Techniques: Inhibition, Activity Assay, Injection, Control, Clinical Proteomics, Muscles

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Association of circulating PLA2G7 levels with cancer cachexia and assessment of darapladib as a therapy

doi: 10.1002/jcsm.12758

Figure Lengend Snippet: Graphical summary of the study. Circulating PLA2G7 levels are increased in CCx in both mice and humans. Despite a significant contribution, tumour is probably not the only responsible for the increase in circulating PLA2G7 levels in CCx. Circulating leucocytes and tissues such as liver, spleen, lymph nodes, or adipose tissue also show increased Pla2g7 expression upon cachexia and may significantly contribute to its circulating levels. In tissues, liver Kupffer cells, lymphocytes, and infiltrating myeloid cells are considered to be the main cell types expressing Pla2g7 (according to publicly available single cell RNA sequencing data on the Tabula Muris website, https://tabula‐muris.ds.czbiohub.org/ ). Tumour‐secreted factors as well as increased circulating levels of pro‐inflammatory cytokines and/or platelet‐activating factor (PAF) may promote PLA2G7 expression and secretion by cells from the host. Chronic treatment with the specific PLA2G7 inhibitor darapladib was not sufficient to improve inflammation and to counteract tissue wasting. Future studies should focus on the potential of PLA2G7 as early biomarker for the diagnosis of CCx.

Article Snippet: CM were diluted with fresh media (1:1, v/v for adipocytes; 3:1, v/v for myotubes) before treatment for 48 h. For darapladib treatment, dimethylsulfoxid (DMSO, vehicle) and 1 μM darapladib (Biorbyt #ORB181231) resuspended in DMSO were freshly added to control or C26 CM before treatment of adipocytes and myotubes.

Techniques: Expressing, RNA Sequencing, Biomarker Discovery