Fig. 1.

Generation of Slc33a1^S113R knock-in mice. (A) The targeting strategy for the Slc33a1 genomic locus. * indicates the S113R mutation. (B) Southern blotting analysis of the Slc33a1 locus using genomic DNA isolated from WT and Slc33a1^wt/mut mice. Samples were digested with either EcoRV (top panel) or NheI (bottom panel), and fragments were detected with 5′ probe or 3′ probe. (C) PCR-based identification of WT and Slc33a1^wt/mut mice. PCR with Slc33a1-specific (F1 and R1) and neo-specific (R2) primers produced two independent products of 477 bp (F1+R1) and 320 bp (F1+R2) for the WT allele and Slc33a1^S113R allele, respectively. (D) PCR-RFLP-based identification of WT and Slc33a1^wt/mut mice. PCR products were digested with HpaII. For WT mice, the digestion products yield only one band (342 bp), and for the Slc33a1^wt/mut mice, the products contain three bands (342, 174 and 166 bp).