Figure 2.

Mouse CAF-1 p150 is required for silencing of pluripotency genes during ESC differentiation. (A) RT-qPCR analysis of expression of three pluripotency genes during EB formation. Results are from three independent experiments. Error bar represents means ± SEM. *P < 0.05, ***P < 0.001 (two-tailed Student's t-tests between p150 WT and each KO line). (B) WB analysis of Oct4 and Nanog during EB formation. Tubulin (bottom) was used as loading control. (C) Hierarchical cluster analysis of differentially expressed genes before (ESC, day 0) and after (EB, day 7) differentiation of p150 WT and KO cells identified by RNA-seq. Results from two independent repeats (rep1 and rep2) are shown. (D) GO analysis of the Group 1 genes identified in C. (E) Representative EGFP fluorescence images of two independent reporter lines of p150 WT and KO ESCs during EB formation. The expression of EGFP is driven by the Oct4 distal enhancer. Scale bar: 400 μM.