Zinc finger homeobox-3 (ZFHX3) is a transcription factor implicated in multiple functions including circadian regulation in the hypothalamus and regulation of signalling pathways in peripheral tissues. We recently demonstrated a novel role for ZFHX3 in whole animal metabolic phenotype, with a missense mutation mouse model similar to a known human mutation, leading to shorter body length, lower body weight, food intake and circulating metabolic hormones, with altered hypothalamic gene expression (1). While the previous work hypothesised that these changes are in the brain, here we aimed to investigate peripheral changes in adipose tissue that may be driven by the Zfhx3^Sci mutation. Objectives: This study  investigated differential gene expression of (a) Zfhx3 in adipose tissues,  (b)  Zfhx3 in a rhythmic and dietary intervention in white adipose tissue (WAT) of wildtype mice, and (c) in adipose tissues of Zfxh3^Sci/+ mice vs wildtype littermates. Methods: Animal experiments were conducted at MRC Harwell and University of Bradford according to the Animals (Scientific Procedures) Act, 1986, under UK Home Office Project Licenses 30/3206, P0D6AA50D and P6165EED1, with local ethical approval. Experiment a: 12 week old female C57B6/N mice were humanely killed and multiple adipose tissues were collected (n = 3-5; inguinal, i-;  gonadal, g-; perirenal, p-; and mesenteric, m-; WAT; and interscapular brown adipose tissue, iBAT). Experiment b: 8 week old male and female C57B6/J mice were housed in a 12 h:12 h light:dark light cycle with access to high fat diet (HFD) or nutrient matched control diet (NMCD) for 6 weeks and tissue iWAT was collected at 4 h intervals over a 24 h diurnal timecourse (n = 5-6). Experiment c: male and female Zfxh3^Sci/+ mice and wildtype littermates (n = 9-12) had ad libitum access to standard rodent chow and water  in a 12 h:12 h light:dark light cycle until 12 months old when they were humanely killed (1). Multiple adipose tissues were collected (iWAT, gWAT, pWAT, mWAT and iBAT) and mRNA extracted followed by qPCR analysis for (a,b) Zfhx3, (b) circadian genes (Per2, Dbp) and adipogenic genes (Ppara, Pparg, Srebp1); and (c) genetic markers of adipogenesis (Pparg, Cebpa, Cebpb, Vegfa, Adipoq), glucose transport (Glut1, Glut4), lipid metabolism (Plin1, Fasn, FAbp4) and thermogenesis (Ucp1, Cidea, Dio2). Statistical comparison was by 2-way ANOVA unless indicated. Results: C57B6/N female mice had significantly different expression of Zfhx3 in visceral adipose depots (one way ANOVA) with differences between gWAT and pWAT vs subcutaneous iWAT approaching significance in post hoc tests (Tukey, p = 0.069 and p = 0.084 respectively).  HFD shifted circadian gene expression (p < 0.0001), and increased expression of Ppara (p <0.05), but Zfhx3 expression did not differ by timepoint. In Zfxh3^Sci/+ mice, very little expression differences were detected in WAT tissues, but in iBAT had significant increased expression of Cebpb, Adipoq, Plin1, Dio2, Ucp1 (p <0.05), Vegfa and Cidea (p < 0.01). Conclusion: Zfhx3 was differentially expressed in visceral and subcutaneous adipose tissue and lower with HFD challenge. Zfhx3^Sci  may be involved in BAT physiology, with increased thermogenic gene expression in these leaner mice.