The first aim of this study was to investigate the effect less-sulfated endothelial HS in correlation to tumor volume. Therefore, the volumes were assessed by comparing Rip1-Tag2 mice bearing floxed Ndst-1 alleles and the TecCre recombinase (RT2 Ndst-1f/f TC+) to mice without the TecCre recombinase (RT2 Ndst-1f/f TC-).

The second aim was to investigate the interaction between HRG-derived peptides by in vitro and in vivo methods. Interaction between HRG330 and HS has previously been demonstrated in a filter-binding assay. However, this method shows limitations in regard to the ability to investigate small peptides, since these cannot be trapped on the membrane. Therefore, a gel-shift assay was conducted to test for possible interaction between the peptides and HS.

The third aim was to conduct the sub-intestinal basket assay in zebrafish. It serves as a convenient method to investigate potential antiangiogenic effects of HRG-derived peptides. If HRG-derived peptides inhibit angiogenesis, zebrafish will develop smaller baskets compared to a control group only injected with a marker.