Figure 1. Generation of PDX1-disrupted PFFs using PDX1-TALEN.

(A) Schematic representation of functional domains of PDX1 and the PDX1-TALEN target site. (B) Magnetic separation-mediated enrichment of PDX1-disrupted PFFs. PFFs were cotransfected with PDX1-targeting TALENs and reporter plasmids, followed by magnetic separation of H-2K^k-expressing cells designed to express both mRFP and eGFP. Fluorescence microscopy examination was performed 1 day after magnetic separation. Magnification, 40×. PDX1-TALEN-driven mutations at endogenous chromosomal sites in magnetic separated PFFs. (C) The T7E1 assay were used to detect PDX1-TALEN-driven mutations. Arrows indicate the expected positions of DNA bands cleaved by T7E1. Mutation frequencies (Indels (%)) were calculated by measuring the band intensities). (D) DNA sequences of the PDX1 wild-type (WT) and mutant clones. The region of the target sequence of the PDX1-TALEN is shown in underlined, deleted bases are indicated by dashes. The column on the right indicates the number of inserted or deleted bases.