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Results: 6

1.
FIGURE 5.

FIGURE 5. From: Receptor Activator of Nuclear Factor ?B Ligand (RANKL) Protein Expression by B Lymphocytes Contributes to Ovariectomy-induced Bone Loss.

Deletion of RANKL from T cells does not alter bone mass. A, RANKL mRNA expression in CD19+ B cells, CD3+ T cells, intact spleen (spl), and L5 vertebra measured by RT-PCR (n = 5–7 mice per group). B, serial BMD of RANKLfl/+ (closed circles), Lck-Cre;RANKLfl/+ (open circles), RANKLfl/fl (closed triangles), and RANKLΔT (open triangles) mice measured up to 7 months of age (n = 4–13 animals per group). The same cohort of animals was used for each age. C, percentage of CD19+ B cells and CD3+ T cells in the bone marrow of 6-month-old RANKLfl/fl and RANKLΔT mice determined by flow cytometry (n = 5–7 mice/group). *, p < 0.05 using Student's t test.

Melda Onal, et al. J Biol Chem. 2012 August 24;287(35):29851-29860.
2.
FIGURE 4.

FIGURE 4. From: Receptor Activator of Nuclear Factor ?B Ligand (RANKL) Protein Expression by B Lymphocytes Contributes to Ovariectomy-induced Bone Loss.

Ovariectomy increases B cell number but not RANKL gene expression. 7-month-old wild-type C57BL/6 mice were sham-operated or ovariectomized and killed after 2 weeks. A, changes in femoral and spinal BMD (n = 13–14 per group). B, RT-PCR analysis of RANKL mRNA in CD19+ B cells isolated from the bone marrow (n = 13–14 mice per group). C and D, percentages of CD19+ B cells and CD19+ B cells that also express RANKL on their surface in the bone marrow (n = 6 mice per group). E, mean intensity of cell surface RANKL protein detected on CD19+ B cells from the bone marrow (n = 6 mice per group). F, amount of sRANKL protein present in bone marrow supernatants normalized to total protein level (n = 9–12 mice per group). *, p < 0.05 using Student's t test.

Melda Onal, et al. J Biol Chem. 2012 August 24;287(35):29851-29860.
3.
FIGURE 2.

FIGURE 2. From: Receptor Activator of Nuclear Factor ?B Ligand (RANKL) Protein Expression by B Lymphocytes Contributes to Ovariectomy-induced Bone Loss.

Deletion of RANKL from B cells attenuates ovariectomy-induced bone loss. 6-month-old female RANKLfl/fl and RANKLΔB mice were either sham-operated (white bars) or ovariectomized (Ovx) (gray bars) and killed 6 weeks later. A, uterine weight per body weight. B, percentage change of body weight. C and D, changes in BMD in the femur and vertebrae during the 6 weeks after surgery. E and F, bone volume over tissue volume (BV/TV) and trabecular thickness (Tb.Th) of cancellous bone measured in lumbar vertebra 4 by μCT. G and H, BV/TV and Tb.Th of femoral cancellous bone measured by μCT. I, cortical thickness (Ct.Th) of the femoral midshaft measured by μCT. J, μCT images of femoral cancellous bone. All values are the mean of 10–17 animals per group. *, p < 0.05, effect of operation within genotype. #, p < 0.05, effect of genotype within operation.

Melda Onal, et al. J Biol Chem. 2012 August 24;287(35):29851-29860.
4.
FIGURE 1.

FIGURE 1. From: Receptor Activator of Nuclear Factor ?B Ligand (RANKL) Protein Expression by B Lymphocytes Contributes to Ovariectomy-induced Bone Loss.

B cell RANKL is required for B cell, but not skeletal, development. A, RANKL mRNA expression in CD19+ B cells and CD3+ T cells in the bone marrow, in intact spleen (spl), and in lumbar vertebrae 5 (L5) (n = 7–17 animals per group). B, serial BMD of RANKLfl/+ (closed circles), CD19-Cre;RANKLfl/+ (open circles), RANKLfl/fl (closed triangles), and RANKLΔB (open triangles) mice determined up to 7 months of age (n = 5–15 animals per group). The same cohort of animals was used for each age. C, percentage of B cells (CD19+), pre-B cells (CD45R/B220+CD43), and T cells (CD3+) in the bone marrow of 6-month-old RANKLfl/fl and RANKLΔB mice determined by flow cytometry (n = 5 samples/group). *, p < 0.05 by Student's t test.

Melda Onal, et al. J Biol Chem. 2012 August 24;287(35):29851-29860.
5.
FIGURE 6.

FIGURE 6. From: Receptor Activator of Nuclear Factor ?B Ligand (RANKL) Protein Expression by B Lymphocytes Contributes to Ovariectomy-induced Bone Loss.

Deletion of RANKL from T cells does not alter ovariectomy-induced bone loss. 6-month-old female RANKLfl/fl and RANKLΔT mice were sham-operated (white bars) or ovariectomized (gray bars) and killed 6 weeks later. A, uterine weight per body weight at time of sacrifice (n = 10–14 animals per group). B and C, percentage of CD19+ B cells and CD3+ T cells in the bone marrow measured by flow cytometry analysis (n = 5 animals per group). D–F, femoral, vertebral, and total body BMD changes during the 6 weeks after surgery (n = 10–17 animals per group). G and H, bone volume over tissue volume (BV/TV) and trabecular thickness of L4 cancellous bone measured by μCT. I, cortical thickness in femoral midshafts measured by μCT. All μCT values are the mean of 10–13 animals/group. *, p < 0.05, effect of operation within genotype.

Melda Onal, et al. J Biol Chem. 2012 August 24;287(35):29851-29860.
6.
FIGURE 3.

FIGURE 3. From: Receptor Activator of Nuclear Factor ?B Ligand (RANKL) Protein Expression by B Lymphocytes Contributes to Ovariectomy-induced Bone Loss.

Deletion of RANKL from B cells blunts ovariectomy-induced osteoclastogenesis. Histological and bone marrow cell counts were performed in the mice described in the legend to Fig. 2. A and B, bone area versus tissue area (BA/TA) and trabecular width (Tb.Wi) measured in L4 vertebra. C and D, osteoclast perimeter per bone perimeter (OcPm/B.Pm) and osteoclast number per bone perimeter (OcN/B.Pm) measured by histomorphometric analysis of lumbar vertebra 1–3 (n = 4–5 animals per group). E, histological sections of lumbar vertebrae stained for TRAP activity (osteoclasts stained red) and toluidine blue (original magnification ×630). F and G, RANKL mRNA levels in sorted CD19+ B cells and CD3+ T cells measured by RT-PCR (n = 7–8 animals/group). H and I, percentage of CD19+ B cells and CD3+ T cells in the bone marrow analyzed by flow cytometry (n = 4 animals per group). *, p < 0.05, effect of operation within genotype. #, p < 0.05, effect of genotype within operation.

Melda Onal, et al. J Biol Chem. 2012 August 24;287(35):29851-29860.

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