Fig. 4.

Knocking down Zeb2-NAT is sufficient to enhance reprogramming of old fibroblasts. Fibroblasts were transfected with the indicated LNA Gapmers and induced to reprogram for 3 weeks. a, b Reprogramming efficiency in adult and old fibroblasts based on quantification of AP⁺ colonies. Student’s t test (two-tailed) statistics, *p < 0.05; error bars represent standard deviation; n=experiments with independent cell cultures. c Proportion of colonies that survived re-plating in medium containing LIF and 2i, without doxycycline; colonies were counted at day 8 after replating. At least ten independent colonies where picked from each condition. Fischer’s exact test was used for statistical analysis. d Immunofluorescence for Nanog and SSEA1 in reprogrammed old fibroblasts transfected with anti-Zeb2-NAT oligonucleotides; the corresponding images stained with DAPI are shown (scale bars, 20 μm). e Histological section of a subcutaneous teratoma containing mesoderm (mes), endoderm (end), and ectoderm (ect) 15 weeks after injection of cells into immune-deficient mice. Magnifications as indicated