Dietary Intervention and Genetic Modulation in Mice

6-week-old male C57BL/6J mice were randomly grouped into 8, 12, 16 and 24 weeks in a cross-sectional study using randomizer.com. In a longitudinal study, 6-8-week-old male Pak2^fl/fl and Pak2^cKO mice were randomly allocated and maintained on high fat high sucrose diet (HFHSD, 45% fat and 20% sucrose) (SDS, 824018) or standard chow (SDS, 801960) for 16 weeks.
Glycyrrhizic acid/Glycyrrhizin (a HMGB1 inhibitor, Sigma-Aldrich, G2137), and Vildagliptin (a DPP4 inhibitor, Sigma-Aldrich, SML2302) were administered to C57BL/6J mice and Pak2^cKO mice ad libitum in drinking water, at an estimated dose of 150 mg/kg/day and 3 mg/kg/day, respectively, along with HFHSD according to individual experimental design (see Results). The water pouch was changed every 3 days. For cardiac specific over-expression of PAK2, adeno-associated virus (AAV9-Pak2) was generated previously (Binder et al., 2019 (link)) using human Pak2 cDNA (Source Bioscience, NCBI Accession # BC069613). The recombinant virus (0.5 × 10¹¹ genomic particles) was administered via tail vein after 8 weeks of HFHSD feeding.