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. Author manuscript; available in PMC: 2010 Feb 27.
Published in final edited form as: Nature. 2009 Aug 9;460(7259):1140–1144. doi: 10.1038/nature08311

Fig. 3. Generation and characterization of 2F-p53KD-iPS cells by p53 downregulation.

Fig. 3

(a) Morphology and GFP fluorescence of 2F-p53KD-iPS cell lines. GFP expression is silenced in clone #6. (b) Alkaline phosphatase staining of 2F-p53KD-iPS cell lines. DAPI was used to visualize cell nuclei. (c) Protein levels of Nanog, Oct4, Sox2, Klf4, c-Myc, p53 in 2F-p53KD-iPS cell lines are shown. α-Tubulin was used as loading control. (d) Embryoid bodies (EBs) of 2F-p53KD-iPS cell clones on day 6 of differentiation. EBs were transferred to gelatinized dishes on day 3 to 5 for further differentiation. On day 14, EBs were subjected to immunofluorescence for α-fetoprotein (AFP)/Foxa2 (endoderm), α-sarcomeric actin/GATA4 (mesoderm) and Tuj1/GFAP (ectoderm). (e) Immunofluorescence of teratoma from 2F-p53KD-iPS cells by antibodies against AFP/Foxa2 (endoderm), α-sarcomeric actinin/Chondroitin (mesoderm), Tuj1/GFAP (ectoderm) showed spontaneous differentiation into all three germ layers. (f) Adult chimeric mice obtained from 2F-p53KD iPS lines (#1 and #6) and non-chimeric mouse in C57BL/6J host blastocysts. (g) As of the date of submission, the mating of offspring from clone #6 chimera to a C57BL/6J female generated 1 agouti pup (blue arrow), that together with PCR analysis (not shown) indicate germ line transmission of the 2F-iPS genome.