Evidence for BRAF mutation and variable levels of microsatellite instability in a syndrome of familial colorectal cancer  Joanne Young, Melissa A. Barker, Lisa A. Simms, Michael D. Walsh, Kelli G. Biden, Daniel Buchanan, Ron Buttenshaw, Vicki L.J. Whitehall, Sven Arnold, Leigh Jackson, Takeshi Kambara, Kevin J. Spring, Mark A. Jenkins, Graeme J. Walker, John L. Hopper, Barbara A. Leggett, Jeremy R. Jass  Clinical Gastroenterology and Hepatology  Volume 3, Issue 3, Pages 254-263 (March 2005) DOI: 10.1016/S1542-3565(04)00673-1 Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 1 Pedigrees for 2 of the MSI-V families. (A) Family J, (B) family B, showing multiple individuals affected with CRC (grey solid shapes) over several generations with tumors displaying divergent MSI subtypes. HP, individual with a hyperplastic polyp. Clinical Gastroenterology and Hepatology 2005 3, 254-263DOI: (10.1016/S1542-3565(04)00673-1) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 2 Examples of significant histology features associated with serrated neoplasia. (A) Serrated architecture in CRC. Moderately differentiated adenocarcinoma with serrated architecture. Cells have relatively abundant apical cytoplasm whereas nuclei tend to be located basally rather than elongated and pseudostratified. A small amount of dirty necrosis is shown, but most luminal space is empty. H&E. (B) Serrated architecture in CRC. Moderately differentiated adenocarcinoma in which intra-epithelial budding has given rise to a serrated architectural contour. Nuclei are round to ovoid, with clearing of chromatin and a prominent nucleolus, a cytologic feature associated with DNA methylation. A small amount of dirty necrosis is shown. H&E. (C) Dirty necrosis. Moderately differentiated adenocarcinoma in which lumen is filled with dense eosinophilic material admixed with necrotic cellular debris (dirty necrosis). H&E. (D) Sessile serrated adenoma. Low-power view of sessile serrated adenoma in which glands show complex architecture but neither the cytoplasmic eosinophilia nor cytologic atypia that are found in traditional serrated adenoma. Smooth muscle proliferation within the lamina propria has entrapped basal glands. H&E. Clinical Gastroenterology and Hepatology 2005 3, 254-263DOI: (10.1016/S1542-3565(04)00673-1) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 3 BRAF Tm curves. (A) Raw data melting curves and (B) negative first derivatives of the raw data melting curves. The mutant sequence (red) has a higher Tm than the wild-type sequence (blue), 85°C and 80°C, respectively, owing to the incorporation of a GC-clamp at the 5′ end of the forward mutant primer E. Template control samples are in black. dF/DT, curves shown in B are first derivatives of those shown in A with respect to time (t). Clinical Gastroenterology and Hepatology 2005 3, 254-263DOI: (10.1016/S1542-3565(04)00673-1) Copyright © 2005 American Gastroenterological Association Terms and Conditions