Figure 7. Inhibition of LSD1 induces energy-expenditure genes in obese adipose tissues.

(a,b) Expression of LSD1 and BHC80 in epididymal WAT from ND (black bars) and HFD- (white bars) fed mice. After a 16-hour fasting period, epididymal WAT was collected, and the total RNA was used for quantitative RT–PCR (a). The expression level of the 36B4 gene was used as the internal control. Protein levels are shown by western blot analysis (b). (c) Expression of LSD1 target genes in ND- (black bars) and HFD- (white bars) fed mice. Values are means±s.d. of four mice, and are shown as fold changes relative to ND-fed mice. **P<0.01 versus ND-fed mice by Student's t-test. (d) Efficient introduction of adenovirus vector into cultured WAT. Adenovirus vector, Ad-CAG-eGFP was introduced into isolated epididymal WAT, and the eGFP expression was analysed by fluorescence microscopy. Scale bar indicates 50 μm. (e,f) Expression of energy-expenditure genes after adenovirus-mediated knockdown of LSD1 gene in cultured WAT ex vivo. Epididymal WAT was dissected from either HFD- (e) or ND-fed mice (f), followed by the infection of adenoviruses, Ad-shLSD1 (red bars) or Ad-sh control (black bars). (g) Efficient introduction of adenoviral vectors into epididymal WAT in vivo. Adenovirus vectors carrying the eGFP gene (Ad-CAG-eGFP) was injected into epididymal WAT after incising the outer coat of mice. Four days later, tissues were isolated for the microscopic analysis. Scale bar indicates 2 mm. (h) Expression of energy-expenditure genes after adenovirus-mediated reduction of LSD1 in epididymal WAT in vivo. Control (black bars) or LSD1 (red bars) shRNA-carrying adenoviruses were directly injected into epididymal WAT of HFD-fed mice 4 days before tissue isolation. Values are means±s.d. of triplicate samples. *P<0.05, **P<0.01 versus control shRNA by Student's t-test.