Figure 7.

Molecular evidence for cell fusion and reprogramming of somatic cells to ESCs. A) RT-PCR for OCT4, Nanog, CD45 and GAPDH was performed on ESCs (lane 1), fused cells (lane 2), or on bone marrow (lane 3). B) To verify cell fusion, fused cells were tested for presence of the CAT gene, which should have been lost gene after Cre recombination. Genomic DNA of the CAT CAGZ bone marrow cells was positive (lane 2), while the fused cells (lane 3) and the water control (lane 1) were negative. C) PCR amplification was performed on the lacZ gene, which was found in the genomic DNA of bone marrow cells from the CAT-CAGZ mouse (lane 2), cDNA of fused cells (lane 3), and in the positive control DNA extracted from the liver of a mouse after infection with adenovirus containing lacZ (lane 4). Water control remained negative (lane 1). D–F) Beating cardiomyocytes differentiated from fused ESCs (D) and those from wild-type ESCs (E) were stained for lacZ. Results further confirmed that fused cells transcribed the lacZ gene. Further, expression of cardiac genes NKx2.5, ANF, MEF2C, β-MHC, and α-MHC in the beating cells was confirmed by PCR (F). Lane 1, positive control of cardiac tissue from a murine heart; lane 2, samples from beating cardiomyocytes generated from beating cardiomyocytes; lane 3, water control.