Fig 7.

Up-regulation of UCP1 in primary brown adipocytes by ApoA-I and D-4F. Brown fat precursor cells were isolated from 6–8-week-old male C57BL/6J mice and cultured for 9 days. The differentiated brown adipocytes were treated with 0.1 μM norepinephrine, 20 μg/ml ApoA-I, 20 μg/ml D-4F for 24 hrs as indicated. The mRNA level of UCP1 was determined by real-time RT-PCR and normalized with the level of β-actin mRNA (A). The protein level of UCP1 was determined by Western blotting and normalized with the protein level of tubulin (B). The differentiated brown adipocytes were serum-starved overnight and then treated with 20 μg/ml ApoA-I, 20 μg/ml D-4F for 10 min. before Western blotting to detect phosphorylated and total AMPK (C). The lower panels for (B) and (C) are semi-quantitative data based on densitometry analysis. The data are shown in mean ± S.D. * indicates P < 0.05 and ** for P < 0.01 as comparison with the control group by Student’s t-test.