Aberrant Methylation of the Eyes Absent 4 Gene in Ulcerative Colitis–Associated Dysplasia  Neal K. Osborn, Hongzhi Zou, Julian R. Molina, Ralf Lesche, Joern Lewin, Cathy Lofton–Day, Kristie K. Klatt, Jonathan J. Harrington, Lawrence J. Burgart, David A. Ahlquist  Clinical Gastroenterology and Hepatology  Volume 4, Issue 2, Pages 212-218 (February 2006) DOI: 10.1016/j.cgh.2005.11.009 Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

Figure 1 Direct bisulphite sequencing of the EYA4 promoter region. Sporadic colon cancer samples (left matrix) compared with normal adjacent tissue (right matrix). Columns represent patient samples, rows represent analyzed CpG positions; numbers refer to the position within the PCR fragment. Methylation percent is reported using a gray-scaled color schema. Numbers on the far right represent P values for differences in methylation between cancer and normal colon for each CpG position. Clinical Gastroenterology and Hepatology 2006 4, 212-218DOI: (10.1016/j.cgh.2005.11.009) Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

Figure 2 MSP. (A) Representative gels from sporadic and CUC tissues (Surv, surveillance group; Remote, matched tissues from CUC neoplasia cases). (B) Representative gels from cell lines. U, unmethylated primers; M, methylated primers. Clinical Gastroenterology and Hepatology 2006 4, 212-218DOI: (10.1016/j.cgh.2005.11.009) Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

Figure 3 Cell line study. (A) Baseline EYA4 methylation and mRNA expression. EYA4 mRNA expression was quantified with reverse-transcription PCR and is shown as the EYA4 to the glyceraldehyde-3-phosphate dehydrogenase ratio on the Y axis. (B) EYA4 mRNA expression quantified by reverse-transcription PCR at baseline and after 4 days of demethylation with a combined Sham trial, DAC, and DAC/TSA regimen. Clinical Gastroenterology and Hepatology 2006 4, 212-218DOI: (10.1016/j.cgh.2005.11.009) Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions