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. 2016 Aug 17;44(22):10960–10973. doi: 10.1093/nar/gkw711

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — vU1s participate in early cell fate decisions. (A) Steady state levels of pluripotent stem cell marker mRNAs, including OCT4, NANOG and SOX2, were measured following transfection of human fibroblasts (NHDF) cells with increasing doses (0.0375, 0.0625, 0.125 and 0.25 μg) of a mixed pool of vU1-expressing plasmids (vU1.2, vU1.3, vU1.8, vU1.13 and vU1.20), by qRT-PCR analysis. Changes in pluripotent mRNA levels (left graph), and vU1 levels (that could be specifically amplified) (right graph), are expressed as fold-difference over levels quantitated in cells transfected with control vector alone, which is set to 1.0. Error bars represent SEM of three independent transfection experiments (Two-way ANOVA analysis; ** = P > 0.05, *** = P > 0.001, **** = P > 0.0001). (B) FACS analysis of NANOG expression in human fibroblasts transfected with decreasing doses of the pooled vU1 plasmids, including 0.5, 0.25 and 0.125 μg. iPSCs and pGEM4 transfected human fibroblast (NDHF-1) cells were used as positive and negative controls, respectively. Histograms represent NANOG fluorescence (black line) compared to isotype control (shaded gray). The % of NANOG positive cells is noted in each histogram.