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Oxidative Medicine and Cellular Longevity
Volume 2017 (2017), Article ID 7308501, 11 pages
Research Article

Mutation Spectrum Induced by 8-Bromoguanine, a Base Damaged by Reactive Brominating Species, in Human Cells

1Department of Tumor Pathology, Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan
2Division of Tumor Pathology, Department of Pathology, Asahikawa Medical University, Asahikawa 078-8510, Japan

Correspondence should be addressed to Kazuya Shinmura

Received 17 May 2017; Revised 23 August 2017; Accepted 6 September 2017; Published 30 September 2017

Academic Editor: Janusz Gebicki

Copyright © 2017 Kazuya Shinmura et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Table S1: Number of mutant clones analyzed in the PCR and gel electrophoresis and in the sequencing in the supF forward mutation assay. Supplementary Table S2: Primers used for PCR amplification of TDG and SMUG1 coding exons. Supplementary Table S3: Mutation spectrum of base substitution mutations at the G position in the supF gene on the shuttle vector plasmid pMY189 replicated in human cells, based on previous reports. Supplementary Figure S1: Proportion of mutation types detected at position 144 of supF on 8BrG-containing pMY189 plasmids, which has an 8BrG residue at position 144 of supF, replicated in H1299 cells. Supplementary Figure S2: Detection of the DNA glycosylase activity of each DNA glycosylase protein on double-stranded oligonucleotides containing the previously reported substrate using a DNA cleavage activity assay. Supplementary Figure S3: Substrates used in the DNA cleavage activity assay. Supplementary Figure S4: Comparison of excisional activities of TDG proteins between thymine mispaired with 8-bromoguanine and thymine mispaired guanine. Supplementary Figure S5: Excision statuses of SMUG1 and TDG proteins for T paired with 8BrG, G, C, or A. Supplementary Figure S6: TDG and SMUG1 protein expression levels in LN428 cells.

  1. Supplementary material