Display Settings:

Items per page

Results: 6

1.
Fig. 2

Fig. 2. From: Wnt-Mediated Regulation of Chondrocyte Maturation: Modulation by TGF-?.

Transforming growth factor-beta (TGF-β), parathyroid hormone related peptide (PTHrP), and BMP-2 regulate Wnt expression in Chondrocytes. Embryonic chick cephalic sternal chondrocyte were cultured for 24 h and were treated with either TGF-β or PTHrP or with BMP-2. After 24 h of treatment, total RNA was extracted from the cultures and gene expressions determined by real-time RT-PCR using chick specific primers. The data shown are representative results of three independent experiments; bars, ±standard error of the mean. The symbol, * represents a significant difference compared to controls with P <0.05.

Yufeng Dong, et al. J Cell Biochem. ;95(5):1057-1068.
2.
Fig. 6

Fig. 6. From: Wnt-Mediated Regulation of Chondrocyte Maturation: Modulation by TGF-?.

TGF-β antagonizes β-Catenin-TCF-4 mediated transcription. Similarly, chondrocytes were transfected with the Topflash reporter (500 ng) and β-Catenin (500 ng) alone or with TCF-4 (500 ng) in the presence of TGF-β (5 ng/ml) and luciferase assay performed after 48 h. Transfection efficiency was determined by co-transfection with pRL vector (Promega) and determining the renilla uniformis luciferase activity (A). Embryonic chick upper sternal chondrocyte were transfected with p3TP-Luc (500 ng) and TCF-4 (500 ng) and either mutant wild-type β-Catenin (500 ng) or control vector after 12 h in culture. Two hours after transfection, standard medium was added with or without TGF-β (5 ng/ml) and the cultures were harvested 48 h later for luciferase assay. Transfection efficiency was determined by co-transfection with pRL vector (Promega) and determining the renilla uniformis luciferase activity. Results are represented as relative luciferase activities obtained by dividing the firefly luciferase by renilla luciferase (B).

Yufeng Dong, et al. J Cell Biochem. ;95(5):1057-1068.
3.
Fig. 5

Fig. 5. From: Wnt-Mediated Regulation of Chondrocyte Maturation: Modulation by TGF-?.

BMP-2 enhances β-Catenin-TCF-4 mediated transcription. Embryonic chick upper sternal chondrocyte were transfected with the full-length type X collagen reporter (ABC-640-Luc; 100 ng) and TCF-4 (500 ng) and/or either mutant (non-degradable) or wild-type β-Catenin (500 ng) after 12 h in culture. Two hours after transfection, standard medium was added with or without BMP-2 (50 ng/ml) (A) or TGF-β (5 ng/ml) (B) and the cultures were harvested 48 h later for luciferase assay. A basic PGL3 vector (10 ng) containing the firefly luciferase gene containing the firefly luciferase gene was used to standardize transfection efficiency. Results are represented as relative luciferase activities obtained by dividing the firefly luciferase by renilla luciferase.

Yufeng Dong, et al. J Cell Biochem. ;95(5):1057-1068.
4.
Fig. 4

Fig. 4. From: Wnt-Mediated Regulation of Chondrocyte Maturation: Modulation by TGF-?.

Stimulation of chondrocyte maturation by Wnt8c is enhanced by BMP-2 and abolished by TGF-β. Embryonic chick upper sternal chondrocyte were infected with either a control RCAS virus, or viruses containing an insert for Wnt8c at the time of plating. After an initial 48 h culture period in medium containing fresh viral supernatant mixed in a 1:1 ratio with plating medium, fresh control medium or medium containing TGF-β (5 ng/ml) or BMP-2 (50 ng/ml) was added. Total RNA was extracted from the cultures 5 days later and gene expressions determined by real-time RT-PCR using chick specific primers for either colX (A) or alkaline phosphatase (B). The data shown are representative results of three independent experiments; bars, ±standard deviation.

Yufeng Dong, et al. J Cell Biochem. ;95(5):1057-1068.
5.
Fig. 1

Fig. 1. From: Wnt-Mediated Regulation of Chondrocyte Maturation: Modulation by TGF-?.

Type X collagen and Wnt gene expression in chick chondrocyte cultures. Chondrocytes were isolated from the cephalic (upper) or caudal (lower) sternum or tibia growth plates of day 15 and day 19 chicken embryos as described in Materials and Methods. Total RNA was prepared from cultures at days 2, 4, and 6. Real-time RT-PCR was carried out using primers for chicken type X collagen, Wnt4, Wnt5a, Wnt7a, Wnt8c, and Wnt9a. The relative expression of colX was examined over time in the different cultures (A). Wnt expression was examined over time in lower sternal chondrocytes (B), upper sternal chondrocytes (C), and growth plate chondrocytes (D). Gene expression was normalized to GAPDH. The data shown are representative results of three independent experiments; bars, ±standard error of the mean.

Yufeng Dong, et al. J Cell Biochem. ;95(5):1057-1068.
6.
Fig. 3

Fig. 3. From: Wnt-Mediated Regulation of Chondrocyte Maturation: Modulation by TGF-?.

β-Catenin and Wnt8c cooperate to stimulate chondrocyte maturation. Embryonic chick upper sternal chondrocyte were transfected with the full-length type X collagen reporter (ABC-640-Luc; 100 ng) and lymphocyte enhancer factor 1 (LEF-1) (500 ng) and/or β-Catenin (500 ng) or control vectors after 12 h in culture. Two hours after transfection, standard medium was added and the cultures were harvested 48 h later for luciferase assay. A basic PGL3 vector (10 ng) containing the firefly luciferase gene was used to standardize transfection efficiency. Results are represented as relative luciferase activities obtained by dividing the firefly luciferase by renilla luciferase (A). Similarly, chondrocytes were transfected with ABC-640-Luc (100 ng) or b2-640-Luc (100 ng) and/or β-Catenin (500 ng) and/or T cell factor (TCF)-4 (500 ng) and luciferase assay performed after 48 h (B). Embryonic chick upper sternal chondrocyte were infected with either a control RCAS virus, or viruses containing inserts for either Wnt8c in RCAS BP (B) and/or β-Catenin in RCAS BP (A) (C,D) at the time of plating. After a 48 h culture period in medium containing fresh viral supernatant mixed in a 1:1 ratio with plating medium, fresh control medium was added. Total RNA was extracted from the cultures 3 days later and gene expressions determined by real-time RT-PCR using chick specific primers for either colX (C) or alkaline phosphatase (D). The data shown are representative results of three independent experiments; bars, ±standard error of the mean. The symbol, * represents a significant difference compared to controls with P <0.05.

Yufeng Dong, et al. J Cell Biochem. ;95(5):1057-1068.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk