Figure 2. Disruption of heparan sulfate in Ndst^CKO mutants.

(A) Tail and lens DNA were collected from the new born Le-Cre;Ndst1^flox/+;Ndst2^KO/KO (designated as Le-Cre;Ndst^flox/+) and the homozygous Le-Cre;Ndst1^flox/flox;Ndst2^KO/KO (designated as Ndst^CKO) pups. Genotyping PCR using primers to specifically detect the Ndst1^flox allele (top band, upper panel), the Ndst1^WT allele (lower band, upper panel) or the Ndst^KO allele (lower panel) confirmed that the complete conversion of the Ndst1^flox allele to the Ndst^KO allele in the lens. (B–G) Although the Le-Cre transgene expression was detected in the E10.5 lens vesicle as shown by the GFP reporter (green fluorescence), heparan sulfate 10E4 staining (red fluorescence) appeared normal until E11.5 when staining was reduced specifically in the anterior Ndst^CKO lens cell basement membranes (arrowhead in E) and was not depleted in the entire Ndst^CKO lens until E14.5 (G). At least ten embryos of each genotype were tested for 10E4 staining.