FIG. 1.

Generation of Nrf2 mutant rats. A, The designed target site of zinc finger nuclease (ZFN) in exon 5 of Nrf2 gene. B, Two lines with an Nrf2 mutation in the ZFN target site, Δ7 (left) and +1 (right). Sequences underlined in (A) were shown. C, Genotyping of Nrf2 Δ7 mutation by electrophoresis. Electrophoresis was performed using PCR products in 4% agarose gel (upper panel). The heterozygous sample of Δ7 mutation and wild-type had both 334 and 364 bp. The polyacrylamide gel electrophoresis-based genotyping analysis was analyzed to distinguish a difference between wild-type (334 bp) and Δ7 mutation (327 bp). Annealing and denaturation formed heteroduplex DNA (*), which migrated slower than homoduplex DNA in 9% acrylamide gel (lower panel). D, Genotyping of Nrf2 +1 mutation by a restriction enzyme, BmgT120I. The PCR product (335 bp) with +1 mutation was digested to 151 and 184 bp by BmgT120I.