Generation and Characterization of Lymphatic and PDPN Knockout Mouse Models

Lymphatic-specific Prox1-EGFP reporter mice (FVB background)61 (link) were purchased from Mutant Mouse Regional Resource Centers, FVB/NCrl mice were purchased from Charles River Laboratories, and C57BL/6 mice were purchased from Jackson Laboratory. Mice with inducible deletion of PDPN (Pdpn^f/f;CagCre) and WT littermates (Pdpn^f/w;CagCre) in mixed background (C57BL/6 and 129/Sv) were generated as previously described49 (link). PDPN deletion was accomplished by administering tamoxifen orally (20 μg each day) from P1 to P6. After weaning, the mice were orally administered 1 mg tamoxifen weekly. The Lgals8 KO mouse strain used for this study was created from embryonic stem cell clone (14305A-F8), obtained from the KOMP Repository (www.komp.org) and generated by Regeneron Pharmaceuticals, Inc62 (link). The Lgals8 coding region deletion was achieved by LacZ (bacterial β-galactosidase) reporter gene replacement of chromosome 13 from 12,440,786 (deletion start) to 12,459,212 (deletion end). Details of the primer sequences and predicted PCR products are available at the Velocigene website (www.velocigene.com/komp/detail/14305). The galectin-8 null status of the KO mice is confirmed by western blotting (Supplementary Fig. 11). The Lgals8 KO mice have no obvious defects in lymphatic vessel development examined by gross morphological analysis.