Fig. 4. (A) Expression profile of the leukemia inhibitory factor (LIF) receptor and gp130 receptor shows that these genes are not highly expressed in human ES or EC cells. (B) RT-PCR confirms the microarray data for ES, EC, and YSC cells. PCRs were performed with TaqMaster Mix (Promega), 1 µl of RT reaction, and 5 pmol of each primer in a total volume of 25 µl. The DNA was denatured at 95˚C for 5 min, then 30 cycles of 95˚C for 30 sec, 50˚C for 30 sec, and 72˚C for 30 sec, followed by 7 min at 72˚C. Samples were run on a 2% agarose gel. Primers used for PCR include: 5' LIFR (GTT AGA CTT AAA AGA GCT GAA GCA CCT AC), 3' LIFR (CCT TTG ATT GTG ACA TTC CGC TGC), 5' GP130 (GAT GTT GAC GTG CAG ACT TGG CTA G), 3′GP130 (GCC TGA AAT TAT TGT GAT TCC ATA AAC), 5' FZD7 (CTC CAG TGT GGA CGC GCT GCG), and 3' FZD7 (CCC TTG CTG CTG TGG CTA AGT CTG). Primers for Pou5F1 and actin are shown in Fig. 3.

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