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. 2012 May 14;7(5):e37070. doi: 10.1371/journal.pone.0037070

Figure 1. Generation of Cul4b flox mice.

Figure 1

(A) Strategy for generation of Cul4b floxed targeting vector. On the top was shown the wild-type allele of Cul4b gene. The targeting vector and targeted allele were shown in the middle and at the bottom, respectively. BamHI (labeled B) and XbaI (labeled X) sites are indicated. Black bars indicating the positions of the probes used in Southern blots are also indicated. (B) Southern blot analysis of genomic DNA isolated from wild-type ES cells and selected ES cell clones. BamHI digested DNA was hybridized with 5′ probe and XbaI digested DNA was hybridized with 3′ probe. WT, wild-type allele; F, flox allele. (C) cDNA sequencing of Cul4b gene from the brain tissue of brain-specific knockout mice (Cul4b flox/Y;Nesin-Cre). As predicted, exon 2 was spliced onto exon 6 after the excision of exons 3–5. (D) Real-time RT-PCR analysis of Cul4b mRNA isolated from brain tissues of wild-type males (Cul4b +/Y;Nestin-Cre +/−), wild-type females (Cul4b+/+;Nestin-Cre +/−), heterozygous females (Cul4b +/flox;Nestin-Cre +/−), and conditional knock-out males (Cul4b flox/Y;Nestin-Cre +/−). (E) Western blot analysis of Cul4b protein isolated from brain tissues of wild-type, heterozygous and conditional knockout mice using an anti-Cul4b antibody. Gapdh was used as a loading control.