Figure 2.

The stable expression of GFPFoxA1 in P19 cells correlates with decreased activity of alkaline phosphatase and increased expression of SSEA-3. (A, B) P19 GFPFoxA1 cells normally express pluripotency-related transcription factors such as Oct4, Nanog, and Sox2. Total RNA or proteins samples were isolated from P19 cells and clone 1, 2, and 3 of P19 GFPFoxA1 cells. The mRNA or protein levels of Oct4, Nanog, and Sox2 were measured by RT-PCR assays (A) or Western blotting (B), respectively. Cyclophilin or β-actin was used as the loading control. (C) P19 cells and the clone 1 cells were stained for alkaline phosphatase. (D) GFPFoxA1 stable expression resulted in significant increase of SSEA-3 in P19 cells. Flow cytometry was used to measure cell populations of SSEA-3 in P19 cells and the clone 1 cells. P19 cells were used as negative control cells.