Figure 4.

The primary cells show self-renewal capacity. (A) FACS analysis of GFP in both primary cell lines shows GFP population. (B) Both primary bulk cells are able to form spheroids in suspension culture, and all the spheroids are GFP positive. (C) The number of spheroids (above 100 μm) from 4×10⁴ bulk primary cells. Assays were performed in triplicate and the error bars indicate the difference in the total number. (D) qRT-PCR analysis of four transcription factors Oct3/4, Sox2, Klf4 and c-Myc expression levels in the indicated samples, which according to miPS-LLCev cell, express high level of Sox2 and Klf4 in spheroid growing cells. Each asterisk shows the significance of P <0.05. Scale bars: 100 μm.