Volume 145, Issue 3, Pages 540-543.e22 (September 2013) Identification of Candidate Oncogenes in Human Colorectal Cancers With Microsatellite Instability  Alexandra E. Gylfe, Johanna Kondelin, Mikko Turunen, Heikki Ristolainen, Riku Katainen, Esa Pitkänen, Eevi Kaasinen, Ville Rantanen, Tomas Tanskanen, Markku Varjosalo, Heli Lehtonen, Kimmo Palin, Minna Taipale, Jussi Taipale, Laura Renkonen–Sinisalo, Heikki Järvinen, Jan Böhm, Jukka–Pekka Mecklin, Ari Ristimäki, Outi Kilpivaara, Sari Tuupanen, Auli Karhu, Pia Vahteristo, Lauri A. Aaltonen  Gastroenterology  Volume 145, Issue 3, Pages 540-543.e22 (September 2013) DOI: 10.1053/j.gastro.2013.05.015 Copyright © 2013 AGA Institute Terms and Conditions

Figure 1 A hot spot mutation in the conserved zinc finger domain of the ZBTB2 protein causes an increase in cell proliferation. (A) The ZBTB2 protein consists of 2 domains; BTB/POZ (bric-a-brac tramtrack broad complex/poxvirus and zinc finger) and a zinc finger domain (1–4). Hot spot mutations are marked in red. (B) The mutational hot spot site in ZBTB2 is highly conservative across species. (C) Increased cell proliferation of mutant ZBTB2 (Arg261Trp) in stable HEK293 cells. Error bars depict the standard error of the mean. Gastroenterology 2013 145, 540-543.e22DOI: (10.1053/j.gastro.2013.05.015) Copyright © 2013 AGA Institute Terms and Conditions

Supplementary Figure 1 (A) The hot spot site (R945) in RANBP2 is conserved across species. (B) Conservation of the mutational hot spot site in PSRC1. The hot spot site was not conserved in mouse and the protein sequence was not conserved in chicken, Xenopus tropicalis, and zebrafish. (C) Gene expression levels of PSRC1 (green), ZBTB2 (purple), and RANBP2 (blue) using real-time PCR. Gene expression is shown for mutation-positive tumors and paired normal colon. Fold change of gene expression is shown on the y-axis. Gastroenterology 2013 145, 540-543.e22DOI: (10.1053/j.gastro.2013.05.015) Copyright © 2013 AGA Institute Terms and Conditions

Supplementary Figure 2 Large-scale analysis of protein interactomes for ZBTB2 and PSRC1. Pie diagrams showing distribution of (A) ZBTB2 and (B) PSRC1 interacting proteins according to their molecular and cellular functions. The top 5 cellular and molecular functions are shown. The –log of the P value was calculated by the Fisher exact test. Heat maps of mass spectrometry data for (C) ZBTB2 and (D) PSRC1. The color schemes represent values that depict the relative abundance of each protein. (E) Inducible expression of ZBTB2 in the Flp-In 293 T-REx cells was verified by Western blot analysis using the indicated antibodies. The C-terminally tagged form of ZBTB2 showed higher stability. Gastroenterology 2013 145, 540-543.e22DOI: (10.1053/j.gastro.2013.05.015) Copyright © 2013 AGA Institute Terms and Conditions

Supplementary Figure 2 Large-scale analysis of protein interactomes for ZBTB2 and PSRC1. Pie diagrams showing distribution of (A) ZBTB2 and (B) PSRC1 interacting proteins according to their molecular and cellular functions. The top 5 cellular and molecular functions are shown. The –log of the P value was calculated by the Fisher exact test. Heat maps of mass spectrometry data for (C) ZBTB2 and (D) PSRC1. The color schemes represent values that depict the relative abundance of each protein. (E) Inducible expression of ZBTB2 in the Flp-In 293 T-REx cells was verified by Western blot analysis using the indicated antibodies. The C-terminally tagged form of ZBTB2 showed higher stability. Gastroenterology 2013 145, 540-543.e22DOI: (10.1053/j.gastro.2013.05.015) Copyright © 2013 AGA Institute Terms and Conditions