Figure 1

Expression of ECATs in male germline stem (GS)cells and ES cells. A. RT-PCR was performed for the number of cycles shown on the right. We categorized ECATs into four groups based on their relative expression levels in GS cells compared to ES cells. To monitor amplification from genomic DNA, we amplified samples in which reverse transcriptase was omitted from the reverse transcription reaction (RT-). As negative controls, water was added instead of cDNA (H2O). NAT1 and Gapdh were used as loading controls. B. Expression of Fbx15 variants examined by RT-PCR. Our previous study using 5' RACE showed that Fbx15 is transcribed from different promoters in ES cells and testes in the mouse (unpublished data). The ES cell-specific variant was not detected in GS cells or adult testis, while the testis-specific variant was weakly detected in ES cells. The two transcripts differ only in exon 1 sequence. The common sequence of the two transcripts was also amplified. NAT1 and Gapdh were used as loading controls.C. Real-time PCR quantification of the expression of Oct3/4 and Sox2 target genes. The expression of each gene was normalized with that of Gapdh. The expression in ES cells was set as 1.0.