Fig 2: Expression of the Pparg gene and its targets are different in ABW and LBW pups. (A) qRT-PCR reveals a reduction in the expression of Rxra and AdipoQ in LBW compared to ABW pups at day 1. (B) Western blot analysis showing the reduction in the expression of Rxrα, Pparγ 1 and 2 and AdipoQ in LBW pups. (C) The rat adiponectin gene promoter showing the position of PPRE elements tested in this study (+1 is the transcription start site). (D,E) ChIP analysis of the occupancy of Pparγ in the adiponectin promoter of ABW (D) and LBW (E) pups. *P<0.05, **P<0.01, two-tailed unpaired Student's t-test.

Fig 3: Deletion of adiponectin from the ArcPomc−/−mice had no effect on their metabolic abnormalities. Body weight comparisons of the male (A) and female (E) mice from 4 weeks to 24 weeks of age. Comparisons of the body composition (B: male and F: female) tissue weights (C: male and G: female) and 24-hour food intake (D: male and H: female) between the ArcPomc−/−:Adipoq+/+ and ArcPomc−/−:Adipoq−/− mice (male, n = 7–11, female, n = 9). (I, J, N, and O) Oral glucose tolerance tests (I: male and N: female) and plasma insulin levels collected at corresponding time points (J: male and O: female). Insulin tolerance tests (K: male and P: female) and HOMA-IR (L: male and Q: female) from the ArcPomc−/−:Adipoq+/+ and ArcPomc−/−:Adipoq−/− mice (male: n = 6–11 and female: n = 9). Glucose levels from 16-hour fasted, 2-h refed, and 4-h refed mice (M, male: n = 6–10, R, female: n = 9). Two-tailed unpaired Student's t-tests were used to compare genotype effects in body composition, tissue weights, 24-hour food intake, HOMA-IR, fasting, and refed glucose levels. Males: 17–22 weeks old; females: 8–12 weeks old.

Fig 4: Rosiglitazone treatment or Wnt10b transgene expression does not alter adiponectin transcript expression in WAT. At necropsy, iWAT and gWAT were dissected, and total RNA was isolated from portions of each tissue. Expression of Adipoq in female (A) and male mice (B) was determined by qPCR and is presented normalized Ppia mRNA expression. Across all groups, statistically significant effects of TZD are presented as described for Figure 2, while significant effects of genotype are described in the text. There were no statistically significant differences between TZD-treated and untreated mice (within each genotype) or between WT and Ocn-Wnt10b mice (within each TZD group), as determined by two-way ANOVA.

Fig 5: Generation of a BMAd-specific Cre mouse model (BMAd-Cre).(A) Efficacy of Osterix-FLPo was evaluated by crossing with FLP-dependent EGFP reporter to yield Osterix-EGFP. (B) Osterix-EGFP male mice at 16 weeks were sacrificed. Fresh tissue confocal microscopy was performed on bisected caudal vertebrae, distal tibia and proximal tibia. Red arrows indicate singly dispersed BMAds. Scale bar; 100 μm. (C) Frozen-sections of proximal tibial slides from Osterix-EGFP were stained with Anti-GFP (green) and DAPI (blue). Scale bar; 50 μm. (D) Schematic of how Osterix-FLPo recombines FLPo-activated Adipoq-Cre (FAC) in BMAd-Cre mice to restrict expression of Cre to BMAds. (E) BMAd-Cre mice were bred with mT/mG reporter mice and the resulting BMAd-mT/mG mice were sacrificed at 16 weeks of age. Cellular fluorescence was evaluated by fresh tissue confocal microscopy. Scale bar; 100 μm. (F) Proximal tibial sections from BMAd-mT/mG mice were stained with antibodies to tdTomato (red) and EGFP (green), and nuclei were counterstained with DAPI (blue). * indicates BMAds. Scale bar; 50 μm.