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. 2019 Oct 10;294(46):17555–17569. doi: 10.1074/jbc.RA119.009409

Figure 2.

Figure 2.

FOXO1 inhibition suppresses chondrogenic differentiation. A, ATDC5 cells were differentiated for 1 week after transfection with siControl or siFOXO1. Levels of FOXO1 protein on days 0, 4, and 7 were determined by Western blotting. Relative mRNA levels of Foxo1, Sox9, Col2a1, Acan, and Col10a1 were measured by qRT-PCR. Gene expression at each stage is expressed relative to the level on day 0 in cells transfected with siControl; n = 4. B, ATDC5 cells were differentiated for 2 weeks after incubation with or without AS1842856 (0.1 μm) for 24 h. Relative mRNA levels of Foxo1, Sox9, Col2a1, Acan, and Col10a1 were measured by qRT-PCR. Gene expression at each stage is expressed relative to the level on day 0 in cells incubated without AS1842856; n = 3. C, ATDC5 cells were differentiated for 4 weeks after incubation with or without AS1842856 (0.1 μm) for 24 h, and Alcian blue staining was performed. D, Alcian blue-stained cultures were extracted and measured at 620 nm; n = 4. Data are presented as mean ± S.D. Statistical analysis was performed using Wilcoxon's rank-sum test. *, p < 0.05.