Figure 1 Locations of CpG dinucleotides within the PGC1α promoter. Locations are defined by bp relative to the transcription start site ( 25 ). Putative transcription factor response elements (underlined) were identified by MatInspector ( http://www.genomatix.de/cgi-bin//matinspector ). AP2, adapt... More

FIG. 3. Liver weight, glycogen content, and gene expression analysis of Hep-M3-KO mice (■) and control littermates (□) maintained on regular diet. A: Liver weight. B: Liver glycogen content of Hep-M3-KO mice and control littermates (freely fed 8-month-old males, n = 6 per group). C: Liver gene expression analysis. Gene expression was studied by real-time qRT-PCR using total hepatic RNA prepared from Hep-M3-KO mice and control littermates (freely fed 3-month-old males). Data from three independent experiments were normalized relative to the expression of cyclophilin A, which served as an internal control. Results are presented as percent change in gene expression in Hep-M3-KO mice relative to control littermates (100%). Acly, ATP citrate lyase; AOX, acyl-CoA oxidase; CPT, carnitine palmitoyltransferase; CREB, cAMP-response element binding protein; FAS, fatty acid synthase; GK, glucokinase; IR, insulin receptor; IRS1, IR substrate 1; PC, pyruvate carboxylase; PGC, PPAR coactivator; PPAR, peroxisome proliferator–activated receptor. FIG. 3. Liver weight, glycogen content, and gene expression analysis of Hep-M3-KO mice (■) and control littermates (□) maintained on regular diet. A: Liver weight. B: Liver glycogen content of Hep-M3-KO mice and control littermates (freely fed 8-month-old males, n = 6 per group). C: Liver gene expression analysis. Gene expression was studied by real-time qRT-PCR using total hepatic RNA prepared from Hep-M3-KO mice and control littermates (freely fed 3-month-old males). Data from three independent experiments were normalized relative to the expression of cyclophilin A, which served as an internal control. Results are presented as percent change in gene expression in Hep-M3-KO mice relative to control littermates (100%). Acly, ATP citrate lyase; AOX, acyl-CoA oxidase; CPT, carnitine palmitoyltransferase; CREB, cAMP-response element binding protein; FAS, fatty acid synthase; GK, glucokinase; IR, insulin receptor; IRS1, IR substrate 1; PC, pyruvate carboxylase; PGC, PPAR coactivator; PPAR, peroxisome proliferator–activated receptor. More

Figure 5 Exercise training increases the androgen receptor activity in iWAT of male mice. A–E: Network motif analysis by Network Analyst (A) on the most upregulated genes (FDR <0.01) with 11 days of voluntary wheel running in male iWAT (Gene Expression Omnibus data set GSE68161). List of genes regulated by AR generated by RegNetwork database (B) and relative pathway enrichment analysis using the Kyoto Encyclopedia of Genes and Genomes (KEGG) (C), GO-BP (D), and GO-CC (E) databases. F: Jaspar analysis for Ucp1 promoter region using Eukaryotic Promoter Database-Swiss Institute of Bioinformatics (SIB) website; androgen response element consensus sequences are represented as red bars. G: Quantitative chromatin immunoprecipitation PCR experiment performed on DNA samples precipitated with antibody against AR after treatment with 10 μmol/L testosterone and 10 μmol/L DHT (n = 3/group). Change in mRNA expression level for Ucp1 (H), Prkaa1 (I), Ppargc1a (J), and Esrra (K) on iWAT after 24 h of 10 mmol/L testosterone treatment (n = 6/group). Data are expressed as mean ± SEM. n.s. indicates no significant difference. *P < 0.05; ***P < 0.001. PPAR, peroxisome proliferator–activated receptor. Figure 5. Exercise training increases the androgen receptor activity in iWAT of male mice. A–E: Network motif analysis by Network Analyst (A) on the most upregulated genes (FDR <0.01) with 11 days of voluntary wheel running in male iWAT (Gene Expression Omnibus data set GSE68161). List of genes regulated by AR generated by RegNetwork database (B) and relative pathway enrichment analysis using the Kyoto Encyclopedia of Genes and Genomes (KEGG) (C), GO-BP (D), and GO-CC (E) databases. F: Jaspar analysis for Ucp1 promoter region using Eukaryotic Promoter Database-Swiss Institute of Bioinformatics (SIB) website; androgen response element consensus sequences are represented as red bars. G: Quantitative chromatin immunoprecipitation PCR experiment performed on DNA samples precipitated with antibody against AR after treatment with 10 μmol/L testosterone and 10 μmol/L DHT (n = 3/group). Change in mRNA expression level for Ucp1 (H), Prkaa1 (I), Ppargc1a (J), and Esrra (K) on iWAT after 24 h of 10 mmol/L testosterone treatment (n = 6/group). Data are expressed as mean ± SEM. n.s. indicates no significant difference. *P < 0.05; ***P < 0.001. PPAR, peroxisome proliferator–activated receptor. More

FIG. 4. Resistance to obesity in Wnt10b-A^y mice at 1 year of age. Female wild-type–A^y and Wnt10b-A^y mice at 12 months of age were random fed or fasted overnight (n = 4). A: Body and adipose depot weights. d-wat, dorsolumbar white adipose tissue; O-WAT, ovarian white adipose tissue; P-WAT, perirenal white adipose tissue. B: Adipocyte area analysis and representative pictures of microscopic fields. C: Primary adipocytes were isolated from ovarian adipose tissue from wild-type–A^y and Wnt10b-A^y mice (n = 8). Gene expression was evaluated by quantitative RT-PCR for the indicated genes. Wnt10b-A^y is shown relative to wild type–A^y (means ± SE). C/EBP, CCAAT/enhancer-binding protein; FABP, fatty acid–binding protein 4; FAS, fatty acid synthase; FSP, fat-specific protein; LPL, lipoprotein lipase; PPAR, peroxisome proliferator–activated receptor. D: Blood glucose and serum proteins from random-fed (n = 4) and fasted (n = 4) mice. E: Quantitative RT-PCR of mRNA from perirenal adipose tissue comparing wild-type–A^y (n = 8) and Wnt10b-A^y (n = 8) mice. Statistical significance was evaluated using Student’s t test. *P < 0.05; **P < 0.001. FIG. 4. Resistance to obesity in Wnt10b-Ay mice at 1 year of age. Female wild-type–Ay and Wnt10b-Ay mice at 12 months of age were random fed or fasted overnight (n = 4). A: Body and adipose depot weights. d-wat, dorsolumbar white adipose tissue; O-WAT, ovarian white adipose tissue; P-WAT, perirenal white adipose tissue. B: Adipocyte area analysis and representative pictures of microscopic fields. C: Primary adipocytes were isolated from ovarian adipose tissue from wild-type–Ay and Wnt10b-Ay mice (n = 8). Gene expression was evaluated by quantitative RT-PCR for the indicated genes. Wnt10b-Ay is shown relative to wild type–Ay (means ± SE). C/EBP, CCAAT/enhancer-binding protein; FABP, fatty acid–binding protein 4; FAS, fatty acid synthase; FSP, fat-specific protein; LPL, lipoprotein lipase; PPAR, peroxisome proliferator–activated receptor. D: Blood glucose and serum proteins from random-fed (n = 4) and fasted (n = 4) mice. E: Quantitative RT-PCR of mRNA from perirenal adipose tissue comparing wild-type–Ay (n = 8) and Wnt10b-Ay (n = 8) mice. Statistical significance was evaluated using Student’s t test. *P < 0.05; **P < 0.001. More

Figure 4 mGPDH modulates podocyte function via RAGE signaling. A: Heat map showing the top 30 up- and downregulated genes in isolated glomeruli from diabetic KO and WT mice. B: Gene ontology analysis of differentially expressed genes and the 10 highest-ranking biological process terms are shown. C: Top 10 ranking terms of KEGG analysis in all significantly upregulated genes. D: Immunofluorescence analysis of RAGE and the podocyte marker synaptopodin in kidneys from diabetic KO and WT mice. E: mRNA expression of genes involved in the RAGE pathway was assessed in isolated glomeruli from diabetic KO and WT mice. F: Differentiated podocytes were transfected with the mGPDH overexpression plasmid with 48 h of HG treatment, and mRNA expression of RAGE pathway-related genes is shown. G–P: Podocytes were knocked down with mGPDH and/or RAGE with their specific siRNA and treated with HG for 48 h. DNA fragmentation (G), immunofluorescence staining of the indicated podocyte proteins (H), mRNA expression of the indicated genes (I), OCR and ECAR (J), mtDNA content (K), MitoTracker Green fluorescence (L), MMP (M), mRNA of OXPHOS genes (N), and Ppargc1a (O) and ROS generation (P) detected by the median fluorescence intensity (MFI) of DCF fluorescence are shown. Scale bars: 20 μm for D and H. n = 3 for A–C and F–P; n = 6 mice/group for D and E. The data are presented as means ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. PPAR, peroxisome proliferator–activated receptor; Si-ctrl, siRNA control; TNF-α, tumor necrosis factor-α. Figure 4. mGPDH modulates podocyte function via RAGE signaling. A: Heat map showing the top 30 up- and downregulated genes in isolated glomeruli from diabetic KO and WT mice. B: Gene ontology analysis of differentially expressed genes and the 10 highest-ranking biological process terms are shown. C: Top 10 ranking terms of KEGG analysis in all significantly upregulated genes. D: Immunofluorescence analysis of RAGE and the podocyte marker synaptopodin in kidneys from diabetic KO and WT mice. E: mRNA expression of genes involved in the RAGE pathway was assessed in isolated glomeruli from diabetic KO and WT mice. F: Differentiated podocytes were transfected with the mGPDH overexpression plasmid with 48 h of HG treatment, and mRNA expression of RAGE pathway-related genes is shown. G–P: Podocytes were knocked down with mGPDH and/or RAGE with their specific siRNA and treated with HG for 48 h. DNA fragmentation (G), immunofluorescence staining of the indicated podocyte proteins (H), mRNA expression of the indicated genes (I), OCR and ECAR (J), mtDNA content (K), MitoTracker Green fluorescence (L), MMP (M), mRNA of OXPHOS genes (N), and Ppargc1a (O) and ROS generation (P) detected by the median fluorescence intensity (MFI) of DCF fluorescence are shown. Scale bars: 20 μm for D and H. n = 3 for A–C and F–P; n = 6 mice/group for D and E. The data are presented as means ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. PPAR, peroxisome proliferator–activated receptor; Si-ctrl, siRNA control; TNF-α, tumor necrosis factor-α. More

Figure 1 Proposed novel cross talk between the PVAT and vascular wall. 4-HNE, 4-hydroxynonenal; PI3K/Akt, phosphoinositide-3 kinase/protein kinase B; PPAR-γ, peroxisome proliferator–activated receptor-γ. Figure 1. Proposed novel cross talk between the PVAT and vascular wall. 4-HNE, 4-hydroxynone... More