Supplementary Figure S1 Generation of Nphp3G2A knock-in mice

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Supplementary Figure S1 Generation of Nphp3G2A knock-in mice Forward primer Reverse primer exon 1 2 3 4 25 26 27 1 kb XbaⅠ XbaⅠ WT Nphp3 allele ATG probe 5´ arm loxP loxP 3´ arm Targeting construct * neor GGCGCC Gly Ala XbaⅠ Nphp3G2A allele (flox) * neor Nphp3G2A allele (after Neo deletion) * b c d Nphp3 genotype WT WT/G2A G2A/G2A 5 kb A T G G G C Met Gly A T G G C C Met Ala WT WT/G2A G2A/G2A 0.5 kb Nphp3G2A/G2A WT allele WT Nphp3G2A allele Supplementary Figure S1 Generation of Nphp3G2A knock-in mice. (a) Targeting strategy to replace the second glycine of Nphp3 to alanine. Alleles produced after homologous recombination and Cre recombination are shown. Neo=neomycin resistance cassette. (b) Verification of correct gene targeting (Southern blot). (c) Sanger sequencing confirmed the homozygous c.5G>C mutation in Nphp3G2A/G2A mice but not in wild-type (WT) mice (arrows). (d) PCR-mediated genotyping of WT, Nphp3+/G2A and Nphp3G2A/G2 mice. Product lengths are 375 bp for WT and 502 bp for knock-in alleles.