FIGURE 4.

COCH mediates BMP4 function to sustain self-renewal of mouse ESCs. A, R1 mouse ESCs stably expressing GFP (GFP) or COCH (COCH-1 and COCH-2) were cultured in N2B27 medium containing LIF only for 5 days, and the self-renewal marker expression was determined by qRT-PCR. R1 mouse ESCs stably expressing GFP (GFP) cells were cultured in N2B27 medium containing LIF + BMP and are shown as the positive control. B, COCH-overexpressing cells as well as GFP stably expressing cells were cultured in N2B27 medium containing LIF only for 5 days and then subjected to AP staining. GFP stably expressing R1 cells cultured in N2B27 medium containing LIF + BMP was used as the positive control. C, Coch knockdown (shCoch-1 and shCoch-2) and control (shLuc) R1 cells were cultured in KSR medium supplemented with LIF or N2B27 medium containing LIF+BMP or LIF only for 5 days, and the self-renewal marker expression was determined by qRT-PCR. Data are shown as mean ± S.E. (n = 3) (*, p < 0.05; **, p < 0.01; ***, p < 0.001).