Glucose meter

Oral glucose tolerance test (OGTT) was performed within one week before the end of treatment. The mice fasted for 16 h with water and were treated with a glucose meter (Roche, Basel, Switzerland) to determine fasting blood glucose.A 2 g/kg/bw glucose solution was rapidly gavaged, and blood glucose values were measured at 30, 60, 90 and 120 min after gavage [18 (link)]. The area under the OGTT curve (AUC) value was calculated after the glucose value was taken from 0 to 120 min by gavage of glucose.Insulin resistance index (HOMA-IR) was calculated as follows [19 ]: $\mathrm{HOMA} - \mathrm{IR} =\left(\mathrm{FPG} \times \mathrm{FINS}\right)/22.5$
In the formula, FPG and FINS represent fasting blood glucose and fasting insulin levels, respectively.

Six-week-old male db/db (C57BLKS/J-lepr^db/lepr^db) mice and their nondiabetic wild-type littermates (WT) were provided by the Model Animal Research Center of Nanjing University (Nanjing, China). The animals were acclimatized to the laboratory before use. At 8 weeks of age, mice were classified into four groups (n = 8 each): WT + Box5 and db/db+Box5 mice were i.p. injected with the Wnt5a inhibitor, Box5 (1 mg/kg body weight), every other day for 12 weeks from 8 weeks of age; WT + Vehicle and db/db+Vehicle mice received 0.9% physiologic saline on the same schedule as the control mice.
Blood glucose levels and body weight were measured every 2 weeks. Blood glucose levels were determined using a glucose meter (Roche, Basel, Switzerland). Twenty-four-hour urine was collected from mice using metabolic cages every 2 weeks. Urinary albumin and creatinine levels were measured using enzyme-linked immunosorbent assay (ELISA) kits (Abcam, Cambridge, USA). The urine albumin excretion rate was expressed as the ratio of albumin to creatinine. All mice were sacrificed at 20 weeks and blood and tissue samples were harvested. Serum creatinine levels were determined using the QuantiChrom Aassay Kit (BioAssay Systems, Hayward, CA, USA). The animal experiments were approved by the Laboratory Animals Ethical Committee of Shanghai Jiao Tong University Affiliated Sixth People’s Hospital.