Figure 1.
Oxygen consumption and energy expenditure.
(A) Whole-body oxygen consumption rate during a 12-hour dark/12-hour light cycle for 30-week-old mice fed a high fat diet (n = 7 for WT; n = 7 for Tg). (B) The average value during the 24-hour period. (C) Energy expenditure during 24-hour period in the experiment. FCBW: fat-corrected body weight = body weight−(mass of subcutaneous and perigonadal white adipose tissue). Data are mean ± SEM. *, p<0.05.
Figure 2.
Metabolic pathways altered in WAT of Tg mice: cell differentiation, lipid synthesis, fatty acid oxidation, and thermogenesis. Fold changes in transcript levels are noted beneath the gene symbols. Atp5g1: ATP synthase, H+ transporting, F1 gamma 1, Bmp: bone morphogenic protein, Cac: carnitine/acylcarnitine translocase, Cox: cytochrome c oxidase, Cpt: carnitine palmitoyl transferase, Dgat: diacylglycerol acyltransferase, Kat: 3-ketoacyl-CoA thiolase, Pgc1a: peroxisome proliferator activated receptor gamma coactivator-1 alpha, Prdm16: PRD1-BF1-RIZ1 homologous domain containing 16, Scd: stearoyl-CoA desaturase.
Figure 3.
Quantitative real-time PCR analysis of the expression of genes in WAT of 30-week-old WT and Tg mice fed a high fat diet, related to (A) fatty acid oxidation and thermogenesis, (B) lipid synthesis, and (C) cell differentiation to white/brown adipocytes. Data were normalized by the amount of 36B4 mRNA and expressed relative to the corresponding value for WAT of WT mice; Data are mean ± SEM (n = 9). *, p<0.05; **, p<0.01. Acc: acetyl-CoA carboxylase, Adrb3: beta-3 adrenergic receptor, Fas: fatty acid synthease, Lpl: lipoprotein lipase, Mcd: malonyl-CoA decarboxylase, Srebp1c: sterol regulatory element binding protein 1c.
Table 1.
Gene expression of WAT in microarray analysis.
Figure 4.
Ectopic expression of UCP1 in WAT of 30-week-old PeriATg mice fed a chow diet. (A) Quantitative real-time PCR analysis of Ucp1 mRNA expression in WAT of WT and Tg mice. Data are mean ± SEM (n = 7). **, p<0.01. (B) Western blot analysis showing UCP1 or CPT1 protein expression in WT and Tg mice. (C) Innunohistochemistry of PeriA or UCP1 in WAT of WT and Tg mice. Original magnification, ×400.
Figure 5.
Alteration in the expression of lipid droplet proteins in vivo. (A) Quantitative real-time PCR analysis of Fsp27 mRNA expression in WAT of WT and Tg mice. Data are mean ± SEM (n = 7). ***, p<0.001. (B) Western blot analysis for FSP27, RIP140 and PeriA in WAT of WT and Tg mice.
Figure 6.
Change of lipid droplet size and protein expression in vitro. (A) Microscopic images of 3T3-L1 adipocytes transfected human PeriA or GFP (control) using adenoviral system plus lipofection methods. (B) Western blot analysis for lipid droplet surface proteins (PeriA and FSP27) and Actin (as an internal control) in 30µg of lysates of day10 3T3-L1 cells treated with adenovirus PeriA (Ad PeriA). PeriA protein content increased dose-dependently with increasing PeriA viral titer in 3T3-L1 cells. Signs in a figure ((−), (+) and (++)) mean the amount of transfected adenovirus (none, single and double quantity). (C) Quantitative real-time PCR analysis of mRNA expression in cultured 3T3-L1 cells (white bar, Ad PeriA(−): black bar, Ad PeriA(++); Data are mean ± SEM (n = 6). *, p<0.05; **, p<0.01; ***, p<0.001). mtTFA: mitochondrial transcription factor A, Nrf1: nuclear respiratory factor 1, Vlcad: very long-chain acyl-CoA dehydrogenase, Lcad: long-chain acyl-CoA dehydrogenase, Mcad: medium-chain acyl-CoA dehydrogenase.