Alb cre mice b6 cg tg alb cre 21mgn j

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 Nf2^flox/flox transgenic mice (with targeted exon 2) was described by Giovannini et al. [41 (link)]. Generation of Cd44^flox/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. Nf2^flox/flox;Alb-Cre; and Cd44-deficient mice were crossed to obtain the desired phenotypes. Nf2^flox/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.

Alb-Cre (B6.Cg-Tg(Alb-cre)21Mgn/J) mice, bought from Jackson Laboratories (https://www.jax.org), were bred with Fur^fl/fl mice to generate hepatocyte-specific Furin-deficient mice (LFurKO) [9 (link)]. RIP-Cre^+/− mice (Tg[Ins2-Cre]23Herr) [23 (link)] were bred with animals in which the exon 4 of the Insr gene was flanked by loxP sites (IR^Lox) (https://www.jax.org/strain/006955; accessed on 6 September 2016), or with Fur^fl/fl mice to generate βIRKO and βFurKO mice, respectively. Mice were backcrossed at least 5 times to a C57Bl6J background as previously described [22 (link)]. All the mice were housed in standard cages on a 12 h day/night cycle and fed a standard rodent chow or HFD (45 kJ % from fat (lard), 20 kJ % from proteins, and 35 kJ % from carbohydrates; Ssniff, Germany) in a conventional animal facility of the KU Leuven. Food and water were provided ad libitum. To avoid confounding effects of estrogens on glucose homeostasis [39 (link),40 (link)], only male mice were included in this study.