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. Author manuscript; available in PMC: 2018 May 1.
Published in final edited form as: Mol Cancer Res. 2017 Feb 1;15(5):507–520. doi: 10.1158/1541-7786.MCR-16-0485

Figure 1. Generation of mutant Idh1 conditional knock-in mouse model.

Figure 1

(a) Schematic diagram of mutant Idh1 conditional knock-in mouse. Floxed stop cassette generates a null allele. Cre-recombinase induces excision of stop cassette and expression of Idh1-R132H. Genotyping primers (1, 2 and 3) are shown. (b) PCR-based genotyping using primers shown in (a). Primer pair (1/2) produces a 263-base pair amplicon when the mutant allele is present. Primer pair (1/3) produces a 329-base pair amplicon when the wildtype allele is present. Only wildtype and heterozygous animals were generated (p< 1.0 × 10−9 (Fisher’s exact test). (c) Sequencing-based validation of the R132H mutation among generated animals. (d) Measurement of D-2HG in wildtype and Idh1LSL:R132H/WT NSCs infected with ad-GFP or ad-Cre recombinase. Measurements reflect six independent wildtype NSC lines and six independent Idh1LSL:R132H/WT NSC lines each treated with ad-GFP or ad-Cre (unpaired Student t-test, p<0.0001, error bars indicate SEM).