Fig 1.
Trp53Cre mice develop thymic lymphomas, but no osteosarcomas.
(A) Representative contact Xrays of 6-month-old male mice with the indicated genotypes. (B) Representative images showing a thymic lymphoma in a 6-month-old Trp53Cre mouse. Quantification of the thymic lymphoma incidence is given below. (C) Genomic PCR amplification of the floxed and recombined Trp53 allele with DNA obtained from the indicated tissues of Trp53fl or Trp53Cre mice. (D) Quantification of body and organ weights in the different groups of mice. Data represent mean ± SD (n ≥ 6).
Fig 2.
Trp53Cre mice display increased trabecular bone mass in the femur diaphysis.
(A) Representative μCT images of femora from 6-month-old male mice with the indicated genotypes. The highlighted regions were separately analyzed. (B) Quantification of the trabecular bone volume per tissue volume (BV/TV) in the mid diaphysis (left) and the distal diaphysis (right) in 6-month-old male mice with the indicated genotypes. (C) Representative μCT images of the femoral midshaft from the same mice. (D) Quantification of cortical thickness (Cort. Th.) and porosity (Cort. Por.) in the same mice. Data represent mean ± SD (n ≥ 6). Asterisks indicate statistically significant differences (*p<0.05, **p<0.005, ***p<0.0005).
Fig 3.
The Trp53Cre phenotype is confirmed by femur histology and gene expression analysis.
(A) Representative images of undecalcified femur sections from 6-month-old male mice with the indicated genotypes. Mineralized bone appears in black. The highlighted regions were separately analyzed. (B) Quantification of BV/TV, trabecular thickness (Tb.Th.) and trabecular number (Tb.N.) in the mid diaphysis (top) or the distal diaphysis (bottom) in 6-month-old male mice with the indicated genotypes. (C) qRT-PCR analysis for expression of the indicated genes in the femur, relative to Gapdh. Data represent mean ± SD (n ≥ 6). Asterisks indicate statistically significant differences (*p<0.05, **p<0.005).
Fig 4.
The Trp53Cre bone phenotype is site-specific.
(A) Representative images of undecalcified spine sections from from 6-month-old male mice with the indicated genotypes. Quantification of BV/TV, Tb.Th. and Tb.N., measured in the indicated region, is shown on the right. (B) Representative images of undecalcified tibia sections from the same mice. Quantification of BV/TV, Tb.Th. and Tb.N., measured in the indicated region, is shown on the right. (C) Representative μCT images of humeri from the same mice. Quantification of BV/TV, Tb.Th. and Tb.N., measured in the indicated region, in shown on the right. (D) Serum concentrations of PINP, CTX, RANKL and OPG in the same groups of mice. Data represent mean ± SD (n ≥ 6). Asterisks indicate statistically significant differences (*p<0.05, ***p<0.0005).
Fig 5.
Trp53Cre bone marrow cells display increased osteogenesis, colony formation and proliferation.
(A) Representative images of cultured bone marrow cells from mice of the indicated genotypes, where mineralized matrix was stained by alizarin red at day 10 of osteogenic differentiation. Photometric quantification is shown on the right. (B) AP (alkaline phosphatase) activity staining of osteogenic colonies in cultured bone marrow cells from mice of the indicated genotypes two weeks after plating at low density. Quantification of the colony numbers is shown on the right. (C) Hematoxylin staining of all colonies in the same cultures. Quantification of the colony numbers is shown on the right. (D) BrdU incorporation assay in undifferentiated bone marrow cells from mice of the indicated genotypes. (E) qRT-PCR analysis for p21 and Casp9 expression, relative to Gapdh, in undifferentiated bone marrow cells from mice of the indicated genotypes. Data represent mean ± SD (n ≥ 6). Asterisks indicate statistically significant differences (*p<0.05, **p<0.005, ***p<0.0005).
Fig 6.
The dental phenotype of Rsk2-deficient mice is not corrected by Runx2-Cre -mediated p53 inactivation.
(A) Representative μCT images of mandibular molars (top) and corresponding grayscale images (bottom) from 6-month-old male mice with the indicated genotypes. Quantification of the visible root area is shown on the right. (B) Hematoxylin (top) and toluidine blue (bottom) staining of molars form the same mice. Quantification of the cellular cementum area is shown on the right. Data represent mean ± SD (n ≥ 6). Asterisks indicate statistically significant differences (*p<0.05, **p<0.005, ***p<0.0005).