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. Author manuscript; available in PMC: 2013 Jul 10.
Published in final edited form as: Oncogene. 2008 Apr 7;27(32):4503–4508. doi: 10.1038/onc.2008.94

Figure 3.

Figure 3

β-catenin and Ki-67 staining in intestines. Intestines at 16 weeks showed β-catenin staining in the adenomas of Rassf1a+/+; Apc+/Min mice (a) with increased nuclear accumulation in adenomas of Rassf1a−/−; Apc+/Min mice (b). Adenomas collected at the time of death from Rassf1a+/+; Apc+/Min (c) and Rassf1a−/−; Apc+/Min (d) mice on tumour watch did not show such clear differences in nuclear β-catenin expression. (e, f) Strong nuclear β-catenin staining was observed in an intestinal adenoma arising from an irradiated Rassf1a−/− mouse. In contrast, no difference in the level of expression or subcellular localization of β-catenin was found in intestines from Rassf1a+/+ (g, i) or Rassf1A−/− (h, j) mice at 2 days or 12 weeks (respectively). Intestines at 16 weeks showed no difference in the number or distribution of Ki-67 positive cells in either the normal epithelium (k, l) or adenomas (m, n) of Rassf1a+/+; Apc+/Min and Rassf1a−/−; Apc+/Min mice, respectively. Immunohistochemistry was performed using anti-β-catenin (1:500 dilution; Sigma-Aldrich, Dorset, UK) or anti-Ki-67 (1:50 dilution; sp6 clone, DCS Diagnostics, Hamburg, Germany). Immunohistochemical signal was detected by secondary biotinylated donkey anti-rabbit antibody (1:500 dilution; Stratech Scientific, Suffolk, UK), followed by Vector ABC tertiary kit (Vector Laboratories, Burlingame, CA) according to the manufacturer’s instructions. All immunohistochemistry was performed on a BondMax machine (Vision Biosystems, Newcastle, UK) according to the manufacturer’s instructions and involved antigen retrieval by BondMax Epitope retrieval solution heated on the machine for 20 minutes. All sections shown are representative. Magnification: x400 (a-f) and x200 (g-n).

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