Figure 4.

A pluripotent state does not occur during conversion from HDFs to CCCAs. HDFs were cultured with STK287794 as in Figure 3 for the indicated number of days. (A) Cells were stained with anti-Nanog and anti-Oct3/4 antibodies, and nuclear staining was performed with DAPI. As a positive control, human iPS cells were also stained. Fluorescence microscopic images are shown (original magnification: ×100). (B) RNA was extracted from the cells and subjected to real-time RT-PCR to evaluate Nanog and Oct3/4 mRNA levels. Values (mean ± SD) are normalized with respect to the β-actin mRNA level in each sample. Relative values to the control were calculated (n = 3 for each group). *** p < 0.001 vs. control group. n.s: not significant.