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. 2002 Jun 11;99(12):8265–8270. doi: 10.1073/pnas.082240999

Figure 1.

Figure 1

CTNNB1 targeting. (A) The upper map shows the targeting construct used to disrupt CTNNB1. The 5′ and 3′ arms were obtained from a human bacterial artificial chromosome library and ligated to a selection cassette flanked by two loxP sites. These loxP sites enabled the efficient removal of the cassette after successful targeting events. The β-catenin minigene encodes a S33Y mutant CTNNB1, and the primer site B was incorporated to facilitate rapid PCR identification of knockout (KO) cells. The lower map shows the regions of CTNNB1 that were targeted by the KO construct. (B) Rapid PCR screening was used to identify clones with successful targeting events at the CTNNB1 (PCR), and targeting events were confirmed by Southern analysis (Southern). The WT and KO alleles are labeled accordingly. (C) CTNNB1 was sequenced in KO clones. Parental HCT116 cells (WT/Δ45) possess both mutant (Δ45) and WT CTNNB1. WT KO clones (−/Δ45) possess only mutant CTNNB1, whereas mutant KO clones (WT/−) have only WT CTNNB1. The first base of codon 45 is labeled with an arrowhead.