Adipose Tissue Differentiation in Mice

All animal studies were approved by the Animal Care and Use Committee of Tohoku University. Male nude mice (BALB/cSlc-nu/nu, purchased from Japan SLC, Inc) were fed standard chow (CE-2, CLEA Japan Inc.) ad libitum in a temperature- and humidity-controlled environment with a 12 h light/12 h dark cycle (08:00–20:00) at constant temperature (23 °C). Transplantation of cultured cells was operated as previously reported^{70 (link)}. 3T3-L1 preadipocytes transduced with empty virus or SETD5 were cultured in differentiation medium for two days. Subsequently, cells were collected by scraping and resuspended in the basal medium containing 1 μg/ml insulin and mixed with 0.25% HydroMatrix Peptide Cell Culture Scaffold (Sigma-Aldrich). Cells (1.5–2 × 10⁶ cells) were injected into the subcutaneous region on the upper or lower back of nude mice at 6 weeks of age. Two weeks after injection, transplanted cells were isolated with skin and subjected to histological analysis.