An emerging concept suggests that an aberrant distributed body fat is closely linked to the occurrence of metabolic abnormalities. Mice deficient in steroid receptor coactivator-3 (SRC-3) are shown to be protected against high-fat diet (HFD) induced obesity but little is known about whether visceral (VAT) and subcutaneous adipose tissue (SCAT) distribute differently in SRC-3(-/-) mice versus SRC-3(+/+) mice. Here we reported that under HFD, fat redistributed between VAT and subcutaneous area of SRC-3(-/-) mice. When VAT/SCAT weight ratio (VAT/SCAT ratio) was calculated, SRC-3(-/-) mice had significantly elevated VAT/SCAT ratio in HFD versus normal diet (ND), while VAT/SCAT ratio was similar in SRC-3(+/+) mice under ND and HFD. Serological changes in SRC-3(-/-) mice paralleled the altered fat distribution. In SRC-3(-/-) mice, assays on gene expression revealed an increase in adipogenesis in VAT versus SCAT and an elevation in thermogenesis and lipolysis in SCAT versus VAT, which could explain the preferential fat accumulation in SRC-3(-/-) VAT. Our results presented in vivo evidence that SRC-3 deficiency could lead to fat redistribution under HFD in mice and provided new clues to researches on the pathogenesis of fat redistribution.