All animal work was approved by the Institutional Animal Care and Use Committee of Huazhong University of Science and Technology. Fifty 10-week-old male C57BL/6J mice were randomly divided into two groups and administered adenoviruses (1 × 109 plaque forming units) (Longchamp et al. 2018) expressing the entire coding sequence of murine asprosin (Ad-Asprosin) or a negative control virus encoding green fluorescent protein (Ad-GFP) in the groin subcutaneous white adipose tissue. Adenoviruses were injected into the center of inguinal scWAT pads within a marked area of approximately 0.5 cm in diameter. The injection site on each side was divided into 10 intensive sites to ensure that the adipose tissue in the labeled area could be overexpressed. After adenovirus injection, the mice were housed under 12 h light:12 h darkness cycle with ad libitum access to food and drinking water in a controlled temperature (23°C ± 1°C) for 2 days. Two days later, we exposed the mice to cold exposure (4°C). In the first 6 h, we measured their rectal temperature using a mouse thermometer at specified time points. After 24 h of acclimation (Huang et al. 2017), indicators related to respiratory metabolism were measured using Comprehensive Lab Animal Monitoring System (CLAMS) for the next 24 h. At day 5, the mice were sacrificed and tissues were collected to further examine the expression of asprosin and histomorphology. Adenovirus was recommended a period of 3–7 days following infection to assess effects on gene expression or physiology (Gomez-Banoy & Lo 2017).

HFD mice were fed with a diet containing 60% calories as fat (D12492, Research Diets, New Brunswick, NJ) for 12 weeks. Other mice used in this study were fed with the standard chow diet (D12450B, Research Diets, New Brunswick, NJ).

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