One touch vita

A glucose solution (20% in 0.9% NaCl) was administered by intraperitoneal (i.p.) injection (100 μl/10 g body weight). Blood glucose was measured at five time points (15, 30, 60, 90, and 120 min after i.p. injection of glucose) during the following 4 h. Blood samples were obtained by a small incision on the paw, and glucose levels were measured by using the blood glucometer One Touch Vita (Johnson & Johnson, NY, USA).

Mice were fasted for 12 h before the experiment. A solution of sterile 0.2 g/ml glucose was freshly prepared in 0.9% sterile saline. Baseline glucose measurements were analysed from tail blood before intraperitoneal (ip) glucose injection (2 mg/g body weight) using the OneTouch Vita (LifeScan, Johnson & Johnson, Milpitas, CA, US) system. Blood glucose measurements were taken using tail blood at 0 min, 15 min, 30 min, 60 min, 90 min and 120 min after injection. Areas under the curve during the insulin tolerance test (AUC-ITT) were calculated using the trapezoidal rule.

Mice were placed in new cages prior to starvation. For GTTs, 12 hr fasted mice were injected intraperitoneal (i.p.) with a solution of sterile glucose (2 g/kg body weight) freshly prepared in 0.9% sterile saline. For ITTs, 6 hr fasted mice were injected i.p. with insulin diluted to 0.08 mU/μl in sterile saline for a final delivery of 0.8 mU/g body weight. Baseline glucose measurements were analyzed from tail blood before i.p. glucose or insulin injection (2 mg/g body weight) using the OneTouch Vita (LifeScan, Johnson & Johnson company) system. Blood glucose measurements were taken from the tail blood at the indicated points.
gWAT morphometry staining gWAT was fixed with Antigenfix (Microm Microtech, France), embedded in paraffin, sectioned, and stained with a hematoxylin and eosin (H&E) solution. Slides (4/group) were scanned with Axio-scan, which allowed the scanning of the entire slide at high resolution. Six pictures of six different areas from 1 to 2 sections per sample were chosen and analyzed with image analyzer software (ImageJ). Total areas of adipocytes were traced manually. The total count ranged from 3275 to 7052 adipocytes per condition. The mean surface area of the adipocytes was calculated using image analyzer software (ImageJ). For each sample, 400–1000 adipocytes were counted.