Fig. S2.

(A) A schematic drawing of the targeted and nontargeted Apc allele. (B) Morphology of Apc-rescued clones nos. 45 and 9. Clones nos. 45 and 9 were derived from RTC nos. 1 and 3, respectively. (C) Southern blotting confirmed successful recombination of the Apc allele in clones nos. 45 and 9. (D) Dosage effect of S33 mutant β-catenin on expression of the canonical Wnt target genes. MEFs and ESCs were treated with Dox at different concentrations for 2 d. Results in (Upper) qRT-PCR analysis and (Lower) RT-PCR analysis are shown. We calculated the ∆∆CT from the CT values of each gene and β-actin in qRT-PCR analysis. Leaky expression level was set to zero for S33 mutant β-catenin. For Ascl2 and Lgr5, values at Dox 0 were set as zero. The average value is shown on the log₂ scale with SD. The previously reported Wnt target genes, Ascl2 and Lgr5, are not significantly affected in ESCs and MEFs by S33 mutant β-catenin expression at different levels. In all conditions, Ascl2 and Lgr5 expression is not detectable in MEFs or ESCs by RT-PCR analysis, respectively. M, marker; N.C., negative control. Related to Fig. 2.