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1.
Figure 1.

Figure 1. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

pK5-Transgene construct and screening techniques. A: Diagram of the K5D-type cyclin construct. B: PCR amplification of DNA extracted from mouse tails. β-globin sequence was amplified resulting in a 450-bp product.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.
2.
Figure 2.

Figure 2. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

Western Blot analysis of cyclin D2, cyclin D3, and pRb family expression in epidermis of transgenic mice. Protein lysates of epidermis were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted to a nitrocellulose membrane. Primary antibodies against cyclin D2, cyclin D3, pRb, p107, and p130 were used for immunoblot analysis. The levels of each protein were quantified with a densitometer.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.
3.
Figure 5.

Figure 5. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

Quantification of epidermal proliferation in transgenic and normal sibling mice. A: The bars indicate the labeling index or the percentage of BrdU incorporation in basal cells from interfollicular epithelia. B: Basal cell hyperplasia. The bars indicate the number of nucleated cells in 200-μm interfollicular epithelia. C: The bars indicate the thickness of whole mouse skin in μm. Gray bars, transgenic mice; white bars, normal siblings.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.
4.
Figure 4.

Figure 4. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

Skin phenotype of K5D3 transgenic mice. Representative paraffin-sections of skin from K5D3 transgenic mice (A) and normal siblings (B) were stained with hematoxylin and eosin. Expression of cyclin D3 in transgenic (C) and wild-type skin (D) was determined with specific antibodies. BrdU incorporation of paraffin sections of transgenic (E) and normal (F) skin was detected with mouse monoclonal anti-BrdU antibody. Antibody binding was detected by secondary antibody conjugated with horseradish peroxidase, and the reaction was developed with diaminobenzidine. Original magnification, ×200.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.
5.
Figure 3.

Figure 3. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

Skin phenotype of K5D2 transgenic mice. Representative paraffin-sections of skin from K5D2 transgenic mice (A) and normal siblings (B) were stained with hematoxylin and eosin. Expression of cyclin D2 in transgenic (C) and wild-type skin (D) was determined with specific antibodies. BrdU incorporation of paraffin sections of transgenic (E) and normal (F) skin was detected with mouse monoclonal anti-BrdU antibody. Antibody binding was detected by secondary antibody conjugated with horseradish peroxidase, and the reaction was developed with diaminobenzidine. Original magnification, ×200.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.
6.
Figure 6.

Figure 6. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

Development of thymic hyperplasia in K5D2 mice. A: Time frame of the development of thymic hyperplasia. Thymus from K5D2 and wild-type mice were harvested and weighed from line 2102 transgenic mice and normal siblings at different ages from 2 to 30 weeks. White bars, transgenic mice; gray bars, normal siblings. B: Thymus of K5D2-2102, K5D3-2203 and normal sibling animals at 11 weeks of age. C: Western blot analysis of cyclin D2 and cyclin D3 expression from protein lysates of thymus of K5D2 (2101 and 2102), K5D3 (2203 and 2201) and normal sibling mice (Wt) at 12 and 24 weeks of age.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.
7.
Figure 7.

Figure 7. From: Cyclin D2 Overexpression in Transgenic Mice Induces Thymic and Epidermal Hyperplasia whereas Cyclin D3 Expression Results Only in Epidermal Hyperplasia.

CDK complex formation and kinase activities in K5D2 thymus. A: Fresh protein lysates of K5D1-2102 transgenic mice thymus at 12 and 20 weeks of age were immunoprecipitated with polyclonal anti-CDK4 (IP/CDK4) and anti-CDK2 (IP/CDK2) antibodies and immunoblotted with polyclonal antibody for p27Kip1, p21Cip1, and cyclin D2. The control was immunoprecipitated with normal rabbit serum with lysates obtained from 2102 transgenic thymus at 12 weeks of age (NR/12). B: CDK4 and CDK2 kinase activities of K5D2 transgenic (T) and normal sibling animals (wt). Fresh protein lysates of thymus were immunoprecipitated as below in an in vitro kinase assay, with pRb as a substrate, was performed. pRb phosphorylation levels were determined by measuring 32P incorporation and Bio-image analysis was used to quantitate the kinase activities. C: Western blot analysis of pRb expression in thymus of transgenic and wild-type mice. Protein lysates of thymus were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted to a nitrocellulose membrane. Primary antibody against pRb was used for immunoblot analysis. T, transgenic; wt, wild type, NR, normal rabbit serum.

Marcelo L. Rodriguez-Puebla, et al. Am J Pathol. 2000 September;157(3):1039-1050.

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