In total, 122 mice aged between 2 and 48 weeks were analyzed to investigate the influence of NF2 and CD44 on different stages of liver tumor development and progression. All animals used in this study were housed under constant temperature and humidity conditions, followed a 12-h light/dark cycle, and had ad libitum access to food and water. All experiments involving mice were approved by the local authorities (AZ: 23 177-07/G 16-1-028). Alb-Cre (B6.Cg-Tg(Alb-cre)21Mgn/J) mice were from The Jackson Laboratory (Sacramento, CA, USA). Generation and genotyping of Nf2flox/flox transgenic mice (with targeted exon 2) was described by Giovannini et al. [41 (link)]. Generation of Cd44flox/flox mice (with targeted exon 3) was described by Dhar et al. (2018) [42 (link)]. Generation and genotyping of Cd44−/− mice was described by Ma et al. (2020) [43 (link)]. All mice were inbred onto C57BL/6 genetic background. Nf2flox/flox;Alb-Cre; and Cd44-deficient mice were crossed to obtain the desired phenotypes. Nf2flox/flox;Alb-Cre; mice were monitored weekly for occurrence of tumors or decline of health, then euthanized after two years at the latest and analyzed.
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