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1.
Fig 2

Fig 2. From: DICKKOPF HOMOLOG 1 MEDIATES ENDOTHELIN-1-STIMULATED NEW BONE FORMATION.

ET-1 increases IL-6 and DKK1 secretion from calvariae. Calvariae were treated with or without 100 nM ET-1 for six days. Media were collected at days two, four, and six days with replacement of fresh media. IL-6 and DKK1 protein concentrations were measured in the conditioned media by ELISA. (* = p<0.05; ** = p<0.01).

Gregory A. Clines, et al. Mol Endocrinol. ;21(2):486-498.
2.
Fig 6

Fig 6. From: DICKKOPF HOMOLOG 1 MEDIATES ENDOTHELIN-1-STIMULATED NEW BONE FORMATION.

ET-1 activates Wnt signaling in osteoblasts. Murine calvarial organ cultures were treated with (B) and without (A) 100 nM ET-1 for seven days. H&E staining was performed. Immunohistochemistry was performed on adjacent sections using an anti-β-catenin antibody. Examples of cell surface (A) and nuclear staining (B) are indicated by arrows.

Gregory A. Clines, et al. Mol Endocrinol. ;21(2):486-498.
3.
Fig 3

Fig 3. From: DICKKOPF HOMOLOG 1 MEDIATES ENDOTHELIN-1-STIMULATED NEW BONE FORMATION.

IL-6 does not mediate the osteoblast-stimulatory action of ET-1. Murine neonatal calvarial organ cultures were treated singly with 100 nM ET-1, 1 μg/ml goat anti-mouse IL-6 neutralizing antibody (αIL-6), 1 μg/ml goat IgG control antibody or in combination for seven days. New bone formation (A) and osteoblast number (B) were determined by standard histomorphometric analyses. (* = p<0.05; ** = p<0.01, *** = p<0.001)

Gregory A. Clines, et al. Mol Endocrinol. ;21(2):486-498.
4.
Fig 5

Fig 5. From: DICKKOPF HOMOLOG 1 MEDIATES ENDOTHELIN-1-STIMULATED NEW BONE FORMATION.

DKK1 neutralizing antibody stimulates new bone formation. Murine neonatal calvarial organ cultures were treated with 100 nM ET-1, 0.1 μg/ml antibody against human DKK1 (αDKK1), or 0.1 μg/ml goat IgG control antibody for seven days and compared. New bone formation (A) and osteoblast number (B) were determined by histomorphometric analyses. No significant differences were found in osteoblast numbers. (* = p<0.05).

Gregory A. Clines, et al. Mol Endocrinol. ;21(2):486-498.
5.
Fig 4

Fig 4. From: DICKKOPF HOMOLOG 1 MEDIATES ENDOTHELIN-1-STIMULATED NEW BONE FORMATION.

DKK1 blocks ET-1-stimulated new bone formation but not basal osteoblast activity. Murine neonatal calvarial organ cultures were treated with 50 ng/ml DKK1, 100 nM ET-1, or both ET-1 and DKK1 (ET+DKK1) in quadruplicate for seven days and compared to control cultures (C). New bone formation (A) and osteoblast number (B) were determined by histomorphometric analyses. (C) Representative calvarial histology is shown. Width between the arrows indicates new bone formation. No significant differences were found in osteoblast numbers. (* = p<0.05; ** = p<0.01).

Gregory A. Clines, et al. Mol Endocrinol. ;21(2):486-498.
6.
Fig 1

Fig 1. From: DICKKOPF HOMOLOG 1 MEDIATES ENDOTHELIN-1-STIMULATED NEW BONE FORMATION.

Real-time RT PCR of primary osteoblast cultures treated with ET-1. Mouse primary osteoblast cultures were treated with or without 100 nM ET-1 in quadruplicate for 1, 3, 6, 24 and 48 hours. Total RNA was isolated and real-time RT PCR performed. Fold mRNA change in ET-1-treated versus control cultures are shown. Hours of ET-1 treatment are indicated on the X-axis and fold changes in mRNA levels are indicated on the Y-axis. The dashed horizontal line is set at a fold change of 1.0, indicating no difference in ET-1-treated versus control culture message. Significant differences in ET-1 versus control treated samples are indicated (* = p<0.05; ** = p<0.01). Dkk1: dickkopf homolog 1; IL6: interleukin 6; IL11: interleukin 11; Ctgf: connective tissue growth factor; Cyr61: cysteine-rich protein 61; Sgk: serum/glucocorticoid regulated kinase; Snai1: snail homolog 1; Rankl: receptor activator of NF-κB ligand; Timp3: tissue inhibitor of metalloproteinase 3; Tgif: TG interacting factor; Dlx3: distal-less homeobox 3; Dlx5: distal-less homeobox 5.

Gregory A. Clines, et al. Mol Endocrinol. ;21(2):486-498.

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