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. 2009 Jul 15;23(14):1625–1630. doi: 10.1101/gad.1809209

Figure 1.

Figure 1.

Generation of a conditional Hdac8 allele. (A) Strategy to generate a conditional Hdac8 allele. Protein domains, genomic structure, targeting vector, and targeted allele are shown. Exon 4 was flanked by loxP sites (red triangles), and the neomycin resistance cassette (Neo), flanked by FRT sites (gray ovals) was removed by crossing to FLPe transgenic mice. NcoI (N) and SacI (S) sites used for Southern blotting are indicated. Primers used for genotyping are depicted with arrows. (B,C) Representative Southern blots of genomic DNA showing correct targeting of the Hdac8 locus in ES cells. Genomic DNA was digested with NcoI and SacI and probed with 5′ and 3′ probes shown in A. (D) Genotyping of wild-type, heterozygous and homozygous null alleles from tail DNA by genomic PCR shows germline transmission and functionality of the conditional allele. (E) Deletion of Hdac8 in mouse embryonic fibroblasts using a tamoxifen-inducible Cre. Hdac8 mRNA levels were detected using primers spanning the deleted exon 4 at the indicated days after adding 4-hydroxy-tamoxifen (4OHT).