Fig. 1 Generation of iPSCs from T. osimensis.

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Presentation transcript:

Fig. 1 Generation of iPSCs from T. osimensis. Generation of iPSCs from T. osimensis. (A) The female T. Osimensis found in the Amami-Oshima Island. Scale bar, 1 cm. (B) Sex is clearly distinguished by the distance between the anus and genitals. Scale bar, 1 cm. (C) Fibroblast cells were propagated from a tail tip in culture. Scale bar, 100 μm. (D) Phase-contrast images of primary colonies and tet-inducible hNANOG-Venus fluorescent images. A strong Venus fluorescent signal was observed in 5f1-1. Scale bar, 100 μm. (E) Dox-independent appearance of iPSC (4f1-1) colonies and their EOS-GFP fluorescence. Arrowheads indicate colonies observed after the withdrawal of Dox. Scale bar, 100 μm. (F) N2B27 3iL contained a B-Raf inhibitor, SB590885, which prevented iPSC (4f1-1) differentiation. Arrowheads indicate the differentiated cells that appeared in the N2B27 2iL (without SB590885) culture condition. Scale bar, 100 μm. (G) Karyotype analysis confirmed the normal chromosome number (2n = 25) in 4f1-1 and 5f1-1. (H) RT-PCR of endogenous expression of undifferentiated marker genes and exogenous genes in all iPSC lines established. (I) Teratoma with three germ layers was confirmed after transplantation of T. osimensis iPSCs (4f1-1 and 5f1-1) into SCID mice. Scale bar, 50 μm. Arata Honda et al. Sci Adv 2017;3:e1602179 Copyright © 2017, The Authors