Fig. 7.

Incorporation of B4 into transplanted nuclei is necessary for pluripotency gene expression in oocytes. (A) ChIP analysis of B4 binding to pluripotency genes in transplanted nuclei. B4 binding to pluripotency gene promoters and to the major satellite region of ES or RA-ES nuclei was analyzed by ChIP 0.5 h, 6 h, and 25 h following transplantation. Error bars represent mean ± SEM. Data are from two experiments. No Ab, control chip performed in the absence of antibody. (B) Incorporation of B4 in transplanted nuclei (control Ab) can be prevented by injection of anti-B4 antibody into the oocyte GV (Lower Row, Center). C2C12 nuclei were transplanted with or without anti-B4 antibody into oocytes expressing B4-GFP. Then B4-GFP labeling of transplanted nuclei was detected by confocal microscopy. The graph on the right shows quantification of the B4-GFP signal. Error bars indicate mean ± SEM (n = 15 nuclei). (C) Pluripotency gene reactivation requires incorporation of B4 into transplanted nuclei. RT-PCR analysis was carried out in conditions depicted in B. Gene reactivation is inhibited in anti-B4 samples at 24 h in both ES and RA-ES transplanted nuclei. Data are from three experiments.