Normal levels of anticoagulant heparan sulfate are not essential for normal hemostasis
J. Clin. Invest. Sassan HajMohammadi, et al. 111:989 doi:10.1172/JCI15809 [Go to this article.]

Figure 1
Disruption of the mouse Hs3st1 locus. (a) Gene-targeting strategy. Exon 8 bridges two BamHI fragments and includes the entire coding sequence (white box) flanked by 5′ and 3′ noncoding sequences (dark gray boxes). The targeting construct replaces exon 8 with a neo^r expression cassette (light gray boxes) in the same transcriptional orientation. The disrupted locus lacks specific BamHI, SpeI, and BglI sites. Restriction sites for producing targeting arms are shown. Also indicated are the locations of 5′ and 3′ probes for Southern blot analysis genotyping and primer sites for PCR analysis genotyping. (b) Genotyping by Southern blotting. Mouse tail genomic DNA was digested with BamHI, then the Hs3st1 locus was detected with the external 5′ probe (shown in a). The wild-type allele generates a 5.6-kb band, whereas the disrupted allele generates a 12.7-kb band due to loss of a BamHI site. (c) 3-OST-1 activity of mouse plasma and tissue homogenates; n = 3 littermates per group. 3-OST-1 activity creates AT-binding sites within HS (producing HS^act) and is determined by incubating tissue homogenate or plasma samples with [³⁵S]HS and PAPS, then measuring the resultant [³⁵S]HS^act by AT-affinity chromatography.