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Haematology (week 16) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: Non-fasted mice are terminally anaesthetised with ketamine/xylazine, and blood is collected into EDTA coated tubes via the retro-orbital sinus. Whole blood is analysed for the following parameters: White and red blood cell counts, mean corpuscular volume, haemoglobin, erythrocyte indices (haematocrit, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red blood cell distribution width), platelet counts and mean platelet volume. All blood samples are analysed with an automatic haematology analyser using volume impedance principle (scilVet Animal Blood Counter, RAB 015 A Ind.F, 01.09.03).

Fluorescence Activated Cell Sorting (week 16) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: FACS analysis is performed on heparinised blood for the following parameters: Percentages of total T cells, CD4+ and CD8+ T cells, NKT cells, NK cells, B cells, Granulocytes and Monocytes are presented relative to the total CD45+ WBC population. Percentages of memory CD4 and CD4+CD25+ Treg cells are presented relative to the total CD4+ T cell population. Percentages of memory CD8 and mature IgD+ B cells are presented relative to the total CD8+ T cell and B cell populations respectively. All samples are analysed on a BD LSR II.