(DOCX 17 KB) Additional file 3: Primers for loss of heterozygosity analysis by single nucleotide polymorphism genotyping. Sequences of the primers used for SNP

LOH PF-01367338 concentration evaluation are shown. All primers designed for use on the Sequenom MassARRAY platform. The percentage of heterozygosity among informative SNPs within two populations from the International HapMap Project are listed. (CEU = Utah residents with Northern and Western European Ancestry; YRI = Samples from Yoruba descent Ibadan, Nigeria; UEP = unextended primer). (DOCX 20 KB) Additional file 4: Characterization of SOSTDC1-specific antiserum. A) A renal cell carcinoma sample with LOH at the SOSTDC1 locus was treated with and without SOSTDC1 antiserum as an internal control to demonstrate effective SOSTDC1 detection. B) Increasing amounts of recombinant SOSTDC1 protein were gel-resolved and immunoblotted with SOSTDC1 antiserum. C) Proteins from the breast carcinoma KU-57788 cell line cell line MDA-MB-231 and those from the breast epithelial cell line MCF10A were resolved and immunoblotted with SOSTDC1-specific antiserum in the presence or absence of competing peptide. The lack of banding in the presence of the immunizing peptide demonstrates

antibody specificity. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein levels were used to verify loading. D) SOSTDC1 was purified from HEK-293 cells transiently transfected to express FLAG epitope-tagged SOSTDC1 protein. The coincident banding when membranes were probed with FLAG-specific antibody and SOSTDC1-directed antiserum verifies the specificity of the antiserum. (TIFF 493 KB) References 1. Aune GJ: Wilms tumor. Pediatr Rev 2008, 29: 142–143. discussion

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