Male black C57BL/6 mice (20 g in weight, 6 weeks of age) were obtained from the National Hellenic Research Foundation and housed under controlled temperature (22±2°C) and photoperiod (12 h light; 12 h dark) with free access to water and food. All experiments with mice were performed in the authorized animal house of the National Hellenic Research Foundation (License no. EL 25 BIObr 031 as a breeding facility; License no. EL 25 BIOsup 032 as a supply facility; and License no. for EL 25 BIOexp 033 as a research installation). The experiments complied with the Protocol on the Protection and Welfare of Animals, as obliged by the rules of the National Hellenic Research Foundation the regulations of the National Bioethics Committee and article 3 of the presidential decree 160/1991 (in line with 86/609/EEC directive) regarding the protection of experimental animals. A total of 3 mice/group were used. The minimum number of animals was achieved according to the 3Rs (replacement, reduction and refinement), to ensure scientific and statistical validity. The health and behaviour of the animals were monitored daily. The mice were gradually euthanized, using carbon dioxide (CO2). The gradual flow rate of CO2 was as per the guidelines. The death of the animals was confirmed through the verification of cardiac and respiratory arrest.

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