Fig.3.

Examining the knock-in ESC clones in terms of zygosity of the Pdx1 locus, subtle mutations in the sgPdx1 target, and random integration of the transgene. A. PCR genotyping of three targeted loci showed that all contained a wild type allele, B. Sequences of sgPdx1 targets in both sides of the targeted allele and the wild type allele contained no mutations and C. Copy number quantification with qPCR demonstrated that all three clones contained only one copy of the GFP sequence when compared with the single copy (hemizygous) Sry locus and the double copy Fgf10 locus. Copy numbers were calculated relative to the mean of Sry. Data are presented as mean ± SD with n=7. K.I.; Targeted knock-in allele, WT; Wild type allele, ESCs; Embryonic stem cells, and qPCR; Quantitative polymerase chain reaction.