recombinant human gdf-5  (PeproTech)

Journal: JCI insight

Article Title: Impaired 1,25-dihydroxyvitamin D3 action underlies enthesopathy development in the Hyp mouse model of X-linked hypophosphatemia.

doi: 10.1172/jci.insight.163259

Figure Lengend Snippet: Figure 8. 1,25D blocks rhGDF5 induced expression of BMP and IHH genes and p-SMAD1/5 and VDR expression is decreased in Hyp entheses by P14. (A) Primary murine chondrocytes were pretreated for 0.5, 1, 4, or 18 hours with or without (–) 1 × 10–8 M 1,25D prior to incubation with (+) or without (–) 100 ng/mL rhGDF5 (for 30 minutes) and subject- ed to Western blot analyses for p-SMAD1/5/9, SMAD1, and β-actin. Data are representative of those obtained from 3 independent experiments. (B) Primary murine chondrocytes were pretreated for 18 hours with 1 × 10–8 M 1,25D followed by treatment with 200 ng/mL rhGDF5 (for 4 hours). Gene expression analyses were performed for BMP and IHH target genes. Data are representative of those obtained from 5–7 independent experiments. One-way ANOVA followed by Fisher’s least significant difference test was used to analyze significance between all genotype groups. *P < 0.05 ver- sus WT; #P < 0.05 versus rhGDF5; a indicates P < 0.05 versus rhGDF5 plus 1,25D. (C) IHC for VDR was performed on P7, P14, P30, and P60 entheses from WT, Hyp, and C–/– mice. In all representative pictures, the enthesis region is outlined with a black box. Scale bar: 20 μm. Data are representative of 6 mice per age or genotype group.

Article Snippet: For Western blot analyses, chondrocytes were pretreated for 0.5, 1, 4, or 18 hours with 1 × 10–8 M 1,25D prior to exposure to recombinant human GDF5 (rhGDF5) (100 ng/mL; R&D Systems) for 30 minutes.

Techniques: Expressing, Incubation, Western Blot, Gene Expression