Canonical Wnt signaling is active in cultured adipocytes and Wntless is upregulated by diet-induced obesity. (A–B) MSCs isolated from C57BL/6J mice were cultured under standard conditions and induced to differentiate. Wntless gene (n = 6) and protein expression at indicated days of differentiation. (C) Wntless gene expression in the stromal-vascular (SVF) and adipocyte (Ads) fractions isolated from epididymal (eWAT) and inguinal (iWAT) white adipose tissues of C57BL/6J mice (males; n = 6). (D) Representative immunoblot of Wntless protein expression across C57BL/6J mouse tissues (asWAT, anterior subcutaneous WAT; pWAT, perirenal WAT; and BAT, brown adipose tissue); adiponectin, laminin, and Ponceau S were included as controls. (E–F) Regulation of Wntless gene and protein expression in eWAT and iWAT of 16-week-old wild-type mice fed a normal chow diet (NCD) or 8 weeks of high-fat diet (HFD) (males; n = 6). (G) Wnt-related genes found to be significantly up- or down-regulated by RNA-seq analyses of day 12 adipocytes vs day 0 cultured MSCs (n = 4). (H) Select Wnt-related genes found to be significantly up- or down-regulated by RNA-seq analyses of day 12 cultured adipocytes treated with recombinant Wnt3a (20 ng/ml) for 4 h (n = 4). RNA expression normalized to PPIA. Data are presented as mean ± S.D. ∗ indicates significance at p < 0.05.