Figure 3. Expression of pluripotency factors in Eed^null ES cells.

Multiple listings for the same gene indicate independent transcripts represented on the microarray and may represent alternate splice forms. Horizontal black lines indicate a value of 1, where mutant and wild-type ES cells have the same expression levels. Dashed black lines represent a value of 0.5, where mutant cells have half of the expression level as wild-type counterparts. (A.) The relative expression levels of several pluripotency markers in low (gray bars) and high (black bars) pass Eed^null ES cells was determined by microarray analysis. While most factors have lower expression levels than wild-type ES cells, gene expression is maintained. With the exception of Klf4, low and high pass mutant cells have similar expression levels. Error bars indicate standard error calculated from 3 technical replicates. (B.) Real-time RT-PCR verification of selected genes represented in panel A shows that expression levels are reproducible. Real-time RT-PCR was carried out in duplicate on RNA generated from pooled cell samples (3 different samples per pool). Error bars represent standard error from 2 replicate pools. (C.) Wild-type and high pass Eed^null ES cells were stained with antibodies against OCT4 and NANOG. Robust staining for is observed for both factors. Corresponding DAPI images are found in Figure S2.