Journal: Development (Cambridge, England)

Article Title: Evolutionarily conserved requirement of Cdx for post-occipital tissue emergence.

doi: 10.1242/dev.079848

Figure Lengend Snippet: Fig. 5. Addition of Fgf8 to the whole embryo culture medium of Cdx2 null embryos rescues their axial elongation. (A,B)E8.0 (early somite) embryos at the start of the culture (A, wild type; B, Cdx2 null). (C-F)Embryos after their culture for the same period without (C,D) or with (E,F) Fgf8 added to the culture medium. C and E are controls; D and F are Cdx2 null mutants. (G-J)Mox1 expression in another set of Cdx2 null mutants (H,J) and controls (G,I) that have been cultured for the same period with (I,J) and without (G,H) Fgf8. (K)Comparison of posterior elongation of Cdx2 null and control embryos cultured without and with Fgf8. y axis, total number of somites generated in culture; bars on the graph represent median values; n, number of embryos; several experimental data are superimposed as one symbol in the graph as they had the same value. (L)Statistical analysis of the axial growth rescue of Cdx mutants by Fgf8 using the Mann-Whitney U test. al, allantois; flb, forelimb bud. Anterior is to the right in A,B and up in C-J. Scale bars: 0.5 mm. See also supplementary material Fig. S3.

Article Snippet: Recombinant Fgf8 (isoform b) was purchased from R&D Systems (423- F8).

Techniques: Embryo Culture, Expressing, Cell Culture, Comparison, Control, Generated, MANN-WHITNEY

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b protects cartilage in DMM mouse model. A. Immunohistochemical analysis of FGF8b expression on the surface of tibial articular cartilage at different time points during osteoarthritis progression, n = 3. Scale bar: 50 μm. B. Endogenous FGF8b expression gradually decreased after DMM surgery. C. Schematic diagram of intra-articular injection of exogenous FGF8b recombinant protein for the treatment of traumatic OA. They received intra-articular injections of either the solvent PBS (10 μl) or exogenous FGF8b recombinant protein (2 μg/10 μl) once per week. D-E. Representative safranin O-fast green-stained sagittal sections of knee joints 8 and 12 weeks after Sham or DMM surgery. The black dashed boxes indicate the magnified regions. Scale bar: low magnification 200 μm, high magnification 50 μm. F-G. OARSI scores for femoral and tibial cartilage in mice treated with PBS or FGF8b 8 and 12 weeks after DMM or Sham surgery, n = 6. MFC: medial femoral condyle; MTP: medial tibial plateau. Data are presented as mean ± 95 % confidence interval. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Immunohistochemical staining, Expressing, Injection, Recombinant, Solvent, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: Continuous administrated FGF8b causes osteophyte formation. A. X-ray images showing osteophyte formation around the knee joint in Sham and DMM surgery groups treated with intra-articular injections of FGF8b. The white arrows indicate the location of osteophytes in the X-ray images. B. Three-dimensional CT reconstruction images displaying extensive osteophyte formation around the knee joint following FGF8b treatment. The red arrows indicate the location of FGF8b-induced osteophytes in the CT images. Scale bar: 1 mm. C. Safranin O-fast green stained sagittal sections of knee joints 8 weeks after DMM surgery. The black dashed lines indicate osteophyte regions near the femoral growth plate. Scale bar: 500 μm. n = 6. D-E. Statistical assessment of osteophyte size and maturity following FGF8b induction in femur. ns: not significant.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b attenuates articular cartilage degeneration in a dose-dependent manner. A-B. Radiographic (X-ray and CT) evaluation of the overall condition of the knee joint in mice treated with 10 μl of different concentrations of FGF8b (once weekly) starting one week after DMM surgery. Scale bar: 1 mm. The white arrows indicate the location of osteophytes in the X-ray images. The red arrows indicate the location of FGF8b-induced osteophytes in the CT images. C. Representative safranin O-fast green stained images of knee joints treated with a gradient of FGF8b concentrations 12 weeks after DMM surgery. The black dashed box indicates the viewing area at high magnification. Scale bar: low magnification 200 μm, high magnification 50 μm. D. OARSI scoring system was used to evaluate the effects of different doses of FGF8b on knee joint cartilage 12 weeks after Sham or DMM surgery. MFC: medial femoral condyle; MTP: medial tibial plateau. n = 6. Data are presented as mean ± 95 % confidence interval. ns: not significant, ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: Intermittent administration of FGF8b protects cartilage in DMM mouse model. A. Mice underwent sham or DMM surgery, and subsequently received intra-articular injections of PBS or FGF8b (2 μg in 10 μl) every four weeks until tissue harvest at 12 weeks for analysis. B-C. X-ray and CT imaging were used to observe the effects of intermittent FGF8b injections on the knee joint. Scale bar: 1 mm. D. Representative safranin O-fast green stained images of knee joints from the Sham or DMM groups treated with intermittent PBS or FGF8b injections. The black dashed boxes indicate the regions magnified. Scale bar: low magnification 200 μm, high magnification 50 μm. E. OARSI scoring system was used to statistically evaluate the degeneration of knee joint cartilage in traumatic OA treated with intermittent FGF8b. n = 6. Data are presented as mean ± 95 % confidence interval. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Imaging, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b promotes anabolism and inhibits catabolism of articular cartilage in mice. A. Adult wild-type mice (12 weeks old) underwent DMM surgery. Intra-articular injection of PBS or FGF8b (2 μg/10 μl) was intermittent administrated. At 4 weeks post-DMM surgery, articular cartilage was extracted, subjected to RNA transcriptome sequencing. n = 4. B. Volcano plot of differentially expressed genes between the PBS or FGF8b-injected groups post-DMM surgery. C. GO enrichment analysis of upregulated genes in the FGF8b-injected group compared to the PBS-injected group post-DMM surgery. D. GO terms of downregulated genes in the FGF8b-injected group compared to the PBS-injected group. E-F. IHC staining of Sox9 and Acan on the surface of articular cartilage in mice treated with intermittent FGF8b. G-H. IHC images of ColⅩ and Mmp13 after FGF8b treatment. I-L. Relative quantitative statistics of gene expression related to cartilage homeostasis. Scale bar: 50 μm. n = 3. ∗∗P < 0.01.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Injection, Sequencing, Immunohistochemistry, Gene Expression

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b prevents human cartilage from IL-1β induced degeneration. A. Safranin O staining revealed increased proteoglycan content in the extracellular matrix of human cartilage sections after FGF8b treatment for Ex vivo OA model. B-D. IHC staining for Acan, Col X, and Mmp13 was performed on human articular cartilage explants treated with IL-1β and FGF8b. Scale bar: 100 μm, n = 3. E-G. Statistical analysis of relative expression of chondro-homeostasis-associated genes in cartilage explants. H. Human primary chondrocytes were treated with varying concentrations of FGF8b for 24 h, followed by western blotting analysis. I-J. The signal intensities of Sox9 and Runx2 were quantitatively analyzed. K. Human primary chondrocytes were treated with IL-1β (10 ng/ml) to induce an OA-like chondrocyte model, along with FGF8b (25 ng/ml) for 24 h. Western blotting was performed to assess the effects of FGF8b on chondrocyte homeostasis-related markers. L-M. Quantitative analysis of Mmp13 and Adamts5 gray value. n = 3. ns: not significant, ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Staining, Ex Vivo, Immunohistochemistry, Expressing, Western Blot

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b exerts a chondroprotective effect via the FGFR3-PI3K-AKT signaling pathway. A. Surface plasmon resonance analysis shows the high-affinity binding of FGF8b to FGFR3. B-C. FGF8b treatment of SW1353 chondrocytes for 15 min increased the expression of phosphorylated FGFR3. D-E. The knockdown efficiency of FGFR3 following transfection with FGFR3 siRNA in SW1353 cells was assessed by Western blotting. F-H. The chondroprotective effect of FGF8b for 24 h was partially attenuated after FGFR3 knockdown. I. KEGG enrichment analysis of transcriptome sequencing data suggested an upregulation of the PI3K-AKT signaling pathway following FGF8b treatment for osteoarthritis. J-L. The FGFR3-PI3K-AKT signaling pathway was downregulated in chondrocytes treated with IL-1β for 15 min, whereas FGF8b treatment activated the FGFR3-PI3K-AKT signaling pathway. M. The activation of the PI3K-AKT signaling pathway by FGF8b is dependent on FGFR3. N-P. The chondroprotective effect of FGF8b was partially abolished upon blockade of the PI3K-AKT signaling pathway using the LY294002 inhibitor under inflammatory conditions. n = 3, ns: not significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 and ∗∗∗∗P < 0.0001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: SPR Assay, Binding Assay, Expressing, Knockdown, Transfection, Western Blot, Sequencing, Activation Assay

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 1. A, B. Arrows show immunostaining of FGF23 in Os and Ob in mouse tibial bone. Scale bars A, B: 50 μm. C. Low magnification image of the areas analyzed: Os (red line) and Ob (blue line). Scale bar: 100 μm. D, E, F. FGF23 expression, visualized by rhodamine IS in Os after 4, 8, 12 days in culture. Scale bars: 100 μm. G. FGF23 WB in Os after 4, 8, 12 days in culture. n = 3–7/ group. Cofilin (bottom of the gel) acted as a loading control. H. qRT PCR of FGF23 mRNA expression in Os. Bars are means ± SD; n = 4–5/group. I. qRT PCR of FGF23 mRNA expression in Ob. Bars are means ± SD; n = 3–5/group. L. FGF23 mRNA expression in Ob and in Os after 12 days in culture. Asterisks indicate significant differences versus Os or Ob T4 (ctrl) (H,I) or Ob T12 (L): ** = p b 0.01, *** = p b 0.001 by ANOVA. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Immunostaining, Expressing, Control, Quantitative RT-PCR

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 3. A. Cultured medium harvested after Ob and Os were cultured for 4, 8, 12 days for measurements of FGF23 protein content, assessed by ELISA. Bars are means ± SD; n = 3/group. B. Cultured medium harvested after Ob and Os were cultured for 4, 8, 12 days for measurements of AHSG protein content, assessed by ELISA. Bars are means ± SD; n = 3/group. Data were normalized by Janus Green Nuclear Marker. Asterisks indicate significant differences versus Os T4 (ctrl): * = p b 0.05, ** = p b 0.01 by ANOVA.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Marker

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 4. A. Changes of AHSG mRNA expression 24 h after the addition of either FGF23 or AHSG to T4 Os. Bars are means ± SD; n = 3–4/group. B. Changes of FGF23 mRNA expres- sion 24 h after the addition of either FGF23 or AHSG to T4 Os. Bars are means ± SD; n = 3/ group. Asterisks indicate significant differences versus Os T4 (ctrl): * = p b 0.05 by ANOVA test.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Expressing

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 5. A. AHSG mRNA expression in Os cultured for 4, 8, 12 days in either control condi- tions or following FGF23 treatment. Bars are means ± SD; n = 3–4/group. B, C. Immuno- fluorescence of AHSG at T4 in control (B) and in FGF23-treated (C) Os. Scale bars: 100 μm. D. WB analysis of AHSG at T4 in control and in FGF23-treated Os; n = 3/group. Cofilin (bottom of the gel) acted as a loading control. E, F. Light microscopy in control (E) and FGF23-treated (F) T4 Os. Scale bars: 100 μm. Asterisks indicate significant differences versus Os T4 (ctrl): ** = p b 0.01 by ANOVA.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Expressing, Cell Culture, Control, Light Microscopy

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 6. A. qRT PCR of FGF23 mRNA in control and FGF23 overexpressing Os at 48 h. Bars are means ± SD; n = 4/group. B. qRT PCR of AHSG mRNA in control and FGF23 overexpressing Os at 48 h. Bars are means ± SD; n = 4/group. C, D. Immunofluorescent detection of AHSG protein at 48 h in control (C) and FGF23 overexpressing (D) Os. Scale bars: 100 μm. E. AHSG WB analysis in control and FGF23 overexpressing Os at 48 h. n = 3/group. Cofilin (bottom of the gel) acted as a loading control. F, G. Light microscopy of control (F) and FGF23 overexpressing Os at 48 h (G). Scale bars: 100 μm. Asterisks indicate significant differences versus Os T4 (ctrl): * = p b 0.05; ** = p b 0.01 Student's t-test.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Quantitative RT-PCR, Control, Light Microscopy

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 7. A. qRT PCR of FGF23 mRNA in control and silenced Os at 48 h after FGF23 gene silencing. Bars are means ± SD; n = 3/group. B. qRT PCR of FGF23 mRNA in control and silenced Os at 48 h after FGF23 gene silencing. Bars are means ± SD; n = 3/group. C, D. Immunofluorescent detection of AHSG protein in control (C) and in FGF23 mRNA silenced Os (D) 48 h after gene silencing. Scale bars C, D: 100 μm. E. AHSG WB in T4 control and FGF23 mRNA silenced Os; n = 3/group. Cofilin (bottom of the gel) acted as a loading control. Asterisks indicate significant differences versus Os T4 (ctrl): ** = p b 0.01 by Student's t-test.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Quantitative RT-PCR, Control

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 8. A. FGFR expression in T4 Os, examined by semi-quantitative PCR. B. qRT PCR of FGF23 mRNA in Os T4 in ctrl, after FGF23 addition, after FGFRs block (FIIN1) addition and after ad- dition of both FGF23 and FGFR blockers. Bars are means ± SD; n = 3–5/group. C. WB analyses of the same (B) experiment n = 3–4/group. Cofilin (bottom of the gel) acted as a loading control. Asterisks indicate significant differences versus Os T4 (ctrl): ** = p b 0.01 by ANOVA.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Blocking Assay, Control

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 9. A. qRT PCR of AHSG mRNA in Os T4 in ctrl, after FGF23 addition, after FGFRs block (FIIN1) addition and after both FGF23 and FGFRs block addition. Bars are means ± SD; n = 4–5/group. B. WB analyses of the same (A) experiment; n = 3–5/group. Cofilin (bottom of the gel) acted as a loading control. Asterisks indicate significant differences versus Os T4 (ctrl): * = p b 0.05, ** = p b 0.01. Triangles significant differences versus FGFRs blocked plus FGF23 treated Os: ▲= p b 0.05 by ANOVA.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Quantitative RT-PCR, Blocking Assay, Control

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 10. A. qRT PCR of AHSG mRNA in control, in FGF23 overexpressing, in FGFR blocked and in both FGFR blocked plus FGF23 overexpressing. Bars are means ± SD; n = 3–4/group. B. WB analyses of the same experiment (A); n = 3/group. Cofilin (bottom of the gel) acted as a loading control. Asterisks indicate significant differences versus Os T4 (ctrl): * = p b 0.05, ** = p b 0.01 by ANOVA.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Quantitative RT-PCR, Control

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 11. A, B. Activation of AHSG promoter detected by secreted Gaussia Luciferase at 24, 36 h after FGF23 overexpression, after addition of FGFR blocker and both FGF23 overexpression and FGFR blocker. Bars are means ± SD; n = 3/group. C. WB analysis of Os T4 detected the FGF23 band in both fractions, the nucleus and the cytoplasm; n = 3/group. Cofilin (bottom of the gel) acted as a loading control. Asterisks indicate significant differences versus Os T4 (ctrl): * = p b 0.05, ** = p b 0.01 by ANOVA.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: Activation Assay, Luciferase, Over Expression, Control

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 12. Immunofluorescence of AHSG (A), FGF23 (B) and merge (C) shows their colocalization. Scale bars: 50 μm.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques:

Journal: Bone

Article Title: FGF23-regulated production of Fetuin-A (AHSG) in osteocytes.

doi: 10.1016/j.bone.2015.10.008

Figure Lengend Snippet: Fig. 13. A, B. Cell localization of AHSG/FGF23 interactions detected by Duolink in situ experiment in the nucleus and in the cytoplasm of Os T4 (A). The negative control exhibits only nuclear DAPI staining showing the presence of cells (B). Scale bars: 50 μm. C. Transformation in white image for quantification with IMAGE J program. Red line indicates total surface area, blue line indicates the nuclear space. Scale bars: 50 μm. D. Quantification of AHSG/FGF23 interactions by following calculation: whole surface area −nucleus area; n = 13 cells. Asterisks indicate significant differences ** = p b 0.01 by Student's t-test.

Article Snippet: 4 day Os were transfected with 10 nM FGF23 mouse cDNA clone (ORF with GFP tagged C terminal) (OriGene Technologies Inc.MD, USA) using Lipofectamine2000 (Invitrogen) as transfecting agent.

Techniques: In Situ, Negative Control, Staining, Transformation 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