Figure 4.

Xanthine oxidase (XO) is induced during adipogenesis, and Febuxostat inhibits ROS-induced excessive adipocyte differentiation. (A) 3T3-L1 and C3H10T1/2 cells were cultured in adipocyte differentiation medium. Xor mRNA expression was determined using real-time PCR. Gapdh served as an endogenous control to normalize each sample. Data are expressed as the mean ± SD (n = 3). * p < 0.05. (B) Xor protein level was analyzed by western blotting. β-actin served as a loading control. Relative changes in the band intensities normalized by respective loading controls are indicated. (C) 3T3-L1 and C3H10T1/2 cells were cultured in an adipocyte differentiation medium with or without N-Acetyl-l-cysteine (NAC) for seven days. Representative image of Oil Red O staining. Original magnification, ×200. Bar, 100 μm. (D) Oil Red O-positive cells were counted. Data are expressed as the mean ± SD (n = 4). * p < 0.05. (E) 3T3-L1 cells were cultured in an adipocyte differentiation medium with the indicated concentration of hydrogen peroxide (H₂O₂) for 48 h. The cells were then washed and further cultured for seven days in the differentiation medium without hydrogen peroxide. Representative image of Oil Red O staining. Original magnification, ×200. Bar, 100 μm. (F) 3T3-L1 and C3H10T1/2 cells were cultured in an adipocyte differentiation medium in the presence or absence of hydrogen peroxide, with or without Febu for 48 h. The cells were then washed and further cultured for seven days in the differentiation medium without hydrogen peroxide with or without Feb. Representative image of Oil Red O staining. Original magnification, ×200. Bar, 100 μm. (G) Oil Red O-positive cells were counted. Data are expressed as the mean ± SD (n = 4). * p < 0.05.