FIGURE 3.

Knockdown of Bcor promotes self-renewal of TSCs. TSCs were infected with lentiviruses carrying control or Bcor shRNAs and selected with puromycin. A, qRT-PCR was performed for Bcor expression in the resultant 129R3 stable cells. B, cells were cultured in 70CM+F4H medium to maintain self-renewal, and the cell numbers were counted every day until day 3. C, cells were cultured in TS medium to induce differentiation, and the cell numbers were counted every the other day until day 5. D, cells cultured in 70CM+F4H medium were subjected to TUNEL assay. Scale bars, 100 μm. The right panel shows the percentage of apoptotic cells counted from five fields of each sample. E, cells were cultured in TS medium plus 25 ng/ml FGF4 and 1 μg/ml heparin (TS+F4H medium) for the indicated times, and the expression of the self-renewal markers Cdx2, Eomes, Elf5, and Esrrb was determined by qRT-PCR. F and G, cells were cultured in TS+F4H medium for 4 days before they were harvested for anti-CDX2 immunostaining (F) or anti-CDX2 immunoblotting (G). GAPDH served as a loading control. Scale bars, 10 μm. Five fields of each sample were quantified for the ratio of CDX2-positive cells. H, cells were cultured in TS medium for the indicated times, and the expression of Cdx2, Eomes, Elf5, and Esrrb was determined by qRT-PCR. Data are presented as mean ± S.E. (n = 3; ***, p < 0.001; **, p < 0.01; *, p < 0.05).