Figure 5: LSD1 demethylase activity is essential for early adipogenesis but dispensable at later stages of differentiation.

(a) Time scale of treatment of primary adipocytes, C3H-10T1/2, or 3T3-L1 cells with LSD1-specific inhibiter GSK690. (b) Western blot analysis of LSD1 and Fabp4 of differentiated primary adipocytes treated with LSD1-specific inhibiter GSK690 or vehicle at the indicated time points of differentiation. β-Tubulin served as a loading control. (c) Relative transcript levels of indicated markers in differentiated primary adipocytes treated with LSD1-specific inhibiter GSK690 or vehicle at the indicated time points of differentiation. (d) Oil red O staining of 3T3-L1, C3H-10T1/2, or differentiated primary adipocytes treated with LSD1-specific inhibiter GSK690 or vehicle at the indicated time points of differentiation. Magnification: × 200; scale bar, 100 μm. (e) Western blot analysis of LSD1 and Fabp4 of (left panel) C3H-10T1/2 or (right panel) 3T3-L1 cells treated with LSD1-specific inhibiter GSK690 or vehicle at the indicated time points of differentiation. β-Tubulin served as a loading control. (f) Relative transcript levels of the indicated markers in C3H-10T1/2 treated with LSD1-specific inhibiter GSK690 or vehicle at the indicated time points of differentiation. (b–f): n=9; s.d. represents+s.e.m. Experiments (c,f) were independently repeated at least three times in triplicate. Statistical analysis was performed using two-tailed Student’s t-test. *P<0.05, **P<0.01 and ***P<0.001.