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. 2009 Jul 3;5(7):e1000547. doi: 10.1371/journal.pgen.1000547

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Gaspar et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Schematic illustration of the chr. 18 LOH event in intestinal tumors from in cis Apc ^+/1572T/Smad4 ^+/Sad mice leading to loss of both Smad4 and Apc wild-type alleles. (B) Comparative phenotypic analysis of the intestinal and mammary tumor predisposition among Apc ^+/1572T, Smad4 ^+/Sad, and Apc ^+/1572T/Smad4 ^+/Sad mice. Notes: (1) The incidence of GI tumors was calculated after exclusion of the pyloric lesions as these present in clusters often difficult to count. (2) The multiplicity of GI tumors was calculated based on all animals with the exception of those where the high tumor burden made the count not feasible. The asterisks indicate that the apparent absence of intestinal tumor in Smad4 ^+/Sad control animals is not in contradiction with what previously published. These mice were sacrificed at time points matched with the ages at which compound Apc ^+/1572T/Smad4 ^+/Sad mice had to be sacrificed due to the high GI and mammary tumor burden (♀: 90.4 days +/−28.4; ♂: 118.5 days +/−26.2). However, in Smad4 ^+/Sad animals the majority of the tumors appear at 9 months of age [15]. (C) H&E staining of intestinal tumor sections from Apc ^+/1572T (top), Apc ^+/1572T/Smad4 ^+/Sad (middle), and Smad4 ^+/Sad (bottom) mice. (D) Smad4 IHC analysis of two intestinal adenomas from Apc ^+/1572T/Smad4 ^+/Sad mice showing loss of Smad4 expression. LOH was observed in 100% of the polyps (n = 15) analyzed. PCR–based LOH analysis of the same cohort of Apc ^+/1572T/Smad4 ^+/Sad polyps revealed loss of wild-type Apc allele in 87% of the cases (13/15; data not shown).