Screening of mutation-restored (Pkd1(+/R+)) iPSCs from iPSCs heterozygous for Pkd1 knockout (KO) (Pkd1(+/−)). a, Normal colony morphology of Pkd1(+/−) iPSCs expressing fluorescence marker protein, GFP. b, Expression of pluripotent marker proteins, Oct4, Sox2, and Nanog in Pkd1(+/R+) iPSCs by immunohistochemical analyses. c, Transcription of pluripotent marker genes in Pkd1(+/−) and Pkd1(+/R+) iPSCs by RT-PCR analyses. d, Secondary screening of Pkd1(+/R+) iPSCs by genomic PCR analyses. e, Verification of replacement of the KO allele by the wild-type (WT) allele through spontaneous mitotic recombination in Pkd1(+/R+) iPSCs by Southern blot hybridization analyses. Relative intensity is noted under the 15.1 kb band. f, Determination of the origin of Pkd1(+/R+) iPSCs by Southern blot hybridization.
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