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2.
Figure 2

Figure 2. p21 protein in Wnt-1 p53+/+ and Wnt-1 p53+/− tumor suspensions.. From: Obesity, Independent of p53 Gene Dosage, Promotes Mammary Tumor Progression and Upregulates the p53 Regulator MicroRNA-504.

A, p21 protein by immunoblot in Wnt-1 p53+/+ and Wnt-1 p53+/− tumor cell suspensions (approximately 1 million cells) and B, in response to UVC DNA damage after 0, 6 and 24 hours. Significant differences in protein levels are indicated by an asterisk on the densitometry plots; P≤0.05.

Nikki A. Ford, et al. PLoS One. 2013;8(6):e68089.
3.
Figure 5

Figure 5. The effect of diet on p53 signaling in Wnt-1 p53+/+ and Wnt-1 p53+/− tumors.. From: Obesity, Independent of p53 Gene Dosage, Promotes Mammary Tumor Progression and Upregulates the p53 Regulator MicroRNA-504.

A, Representative photomicrographs of immunohistochemical staining of Wnt-1 p53+/+ or Wnt-1 p53+/− mammary tumors for p53 and p21 (20×) with B, bar graphs presenting Aperio quantitation from mice fed a control diet or DIO regimen, means ± SD, (n = 5 per group). Significant differences are indicated by an asterisk; P≤0.05. p21 IHC data was natural log transformed to meet statistical test assumptions.

Nikki A. Ford, et al. PLoS One. 2013;8(6):e68089.
4.
Figure 3

Figure 3. Effect of p53 gene dosage and a DIO regimen versus a control diet on Wnt-1 mammary tumor growth.. From: Obesity, Independent of p53 Gene Dosage, Promotes Mammary Tumor Progression and Upregulates the p53 Regulator MicroRNA-504.

A, the percentage of mice with palpable Wnt-1 mammary tumors after tumor cell injection in response to a control diet or DIO regimen, (n = 20 per group). Wnt-1 p53+/+ and Wnt-1 p53+/− final tumor weights are shown in B and C, respectively, and Wnt-1 p53+/+ and Wnt-1 p53+/− final tumor volumes are shown in D and E, respectively (means indicated by horizontal lines). Significant differences are indicated by an asterisk; P≤0.05.

Nikki A. Ford, et al. PLoS One. 2013;8(6):e68089.
5.
Figure 4

Figure 4. Effect of p53 gene dosage and a DIO regimen versus a control diet on Wnt-1 mammary tumor pathology and proliferation.. From: Obesity, Independent of p53 Gene Dosage, Promotes Mammary Tumor Progression and Upregulates the p53 Regulator MicroRNA-504.

A, representative photomicrographs of hematoxylin- and eosin-stained sections of Wnt-1 p53+/+ and Wnt-1 p53+/− tumors from mice fed a control diet or DIO regimen (n = 11–20 mice/group; 10–20×). Each tumor-bearing mouse developed a single tumor. Arrows point to indicated pathological structures. B, representative photomicrographs of immunohistochemical staining of tumors for Ki-67 (20×) and a bar graph presenting the Aperio image quantitation, means ± SD, (n = 5 per group). Significant differences are indicated by an asterisk; P≤0.05.

Nikki A. Ford, et al. PLoS One. 2013;8(6):e68089.
6.
Figure 6

Figure 6. Effect of p53 gene dosage and a DIO regimen versus a control diet on Wnt-1 mammary tumor gene expression of p53 and its regulators.. From: Obesity, Independent of p53 Gene Dosage, Promotes Mammary Tumor Progression and Upregulates the p53 Regulator MicroRNA-504.

mRNA expression, measured by quantitative real-time PCR, of p53, mouse double minute (MDM2), Sirtuin (Sirt)1, microRNA (miR)-125b and miR-504 (n = 5 per gene per group; 3 replicates) in Wnt-1 p53+/+ and Wnt-1 p53+/− tumor tissue from DIO or control mice. Data are presented as gene expression relative to that in Wnt-1 p53+/+ tumors from control mice (means ± SD). Significant differences are indicated by an asterisk; P≤0.05. miR-125b data was natural log transformed to meet statistical test assumptions.

Nikki A. Ford, et al. PLoS One. 2013;8(6):e68089.
7.
Figure 7

Figure 7. Effect of p53 gene dosage and a DIO regimen versus a control diet on Wnt-1 mammary tumor on invasive markers and ERα expression.. From: Obesity, Independent of p53 Gene Dosage, Promotes Mammary Tumor Progression and Upregulates the p53 Regulator MicroRNA-504.

A, representative hematoxylin and eosin images (n = 11–20; 10–20×) of Wnt-1 p53+/+ and Wnt-1 p53+/− tumors and surrounding mammary fat pads from mice fed a control diet or DIO regimen. Representative micrographs of immunohistochemical staining of tumors for e-cadherin, slug, keratin 8 and ERα (20×),B, bar graphs presenting Aperio quantification, and C, bar graphs depicting gene expression of EMT markers, means ± SD, (n = 5 per group). Significant differences are indicated by an asterisk; P≤0.05. Positive staining for ERα and slug was further stratified by 1+, low; 2+, moderate; and 3+, intense staining.

Nikki A. Ford, et al. PLoS One. 2013;8(6):e68089.

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