Glucose tolerance test (GTT) and insulin tolerance test (ITT) in all groups were performed largely as described58 (link). Briefly, after overnight and 5-h fasting, GTT and ITT were conducted at ZT2 and ZT 8 respectively. Glucose levels were measured from tail blood before and 15, 30, 60, or 120 min after i.p. injection of either 1 g/kg glucose or 0.75 U/kg insulin (Eli Lilly) by using the ONETOUCH glucometer (LifeScan).
For insulin challenge, mice were given vehicle (PBS) or insulin (5 U/kg) via i.p following overnight fasting (15–18 h). Five minutes after injection, mice were deeply anesthetized by isoflurane inhalation. Isolated liver samples were immediately frozen in liquid nitrogen and stored in −80 °C freezer until later use.
For insulin challenge, mice were given vehicle (PBS) or insulin (5 U/kg) via i.p following overnight fasting (15–18 h). Five minutes after injection, mice were deeply anesthetized by isoflurane inhalation. Isolated liver samples were immediately frozen in liquid nitrogen and stored in −80 °C freezer until later use.
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