Fig. 3.

Expression of the four reprogramming factors and evidence for reprogramming in mouse liver. a Detection of AAV-encoded hCO-Oct-3/4 (brown staining) via immunohistochemistry in liver sections obtained from mice 2 or 4 weeks post-injection with 5 × 10¹⁰ (low dose) or 2 × 10¹¹ (high dose) vg per vector and mouse (n = 3 per dose and time point). Symbols are the same as in b. Nuclei were counterstained with hematoxylin. Scale bar = 100 µm. b Quantification of the expression of AAV-encoded hCO-Oct-3/4, -Klf4, -Sox2 and -c-Myc by qRT-PCR. A delta Ct analysis was used in which the values were normalized to the expression of a housekeeper gene (actin) and then expressed as fold-changes over the values (set to 1) for the low dose at 2 weeks for each reprogramming factor. Each point represents a single mouse. Center values are means and error bars are S.D. Statistical analyses were performed by one-way ANOVA with Tukey’s Multiple Comparison Test. *, p < 0.05; **, p < 0.01. c, d Evidence for reprogramming in livers of AAV-OKSM-treated mice (2 × 10¹¹ vg, sacrificed 8 weeks after administration) obtained via immunohistochemical detection of expression of Tfe3 (c), a marker for pluripotency, and PCNA (d), a marker for proliferation. Note the largely overlapping patches of Tfe3- and PCNA-positive cells in the serial sections from the same liver of an AAV-OKSM-treated mouse in the rightmost panels. Scale bars in the left and central panels = 100 µm, and in the right panels = 500 µm