NextSeq 500 sequencing; Diet induced obesity is independent of metabolic endotoxemia, Tlr4 signalling and hypothalamic inflammation
Diet induced obesity is independent of metabolic endotoxemia, Tlr4 signalling and hypothalamic inflammation
Mice culled by anaesthetizing with isofluorane followed by cervical dislocation. Brain removed by cutting coronally at approximately bregma -0.1 and -3.0. The slice was orientated with the caudal side face up and a hypothalamic punch of 3 mm in diameter was taken from the ventral aspect. Punches were transferred to a microfuge tube immediately and frozen on dry ice. Tissue was stored at -70 degrees Celsius until processed.  Sixteen wild type C57BL/6J mice, 3-4 weeks old (Harlan), singly housed and acclimatized (12:12 light:dark) on low fat diet (10% fat) for two weeks. Eight mice then switched to high fat diet (60% fat) and eight mice remain on low fat diet (10% fat). Diets continued for 20 weeks. RNA was extracted from the whole hypothalamic block using Machery-Nagel RNA isolation kit as per manufacturer's instructions. Libraries were prepared for sequencing using the Illumina TruSeq Stranded mRNA kit.
Experimental Factor: diet
low fat diet