10.1371/journal.pone.0017557.g003 Junying Yu Junying Yu Kevin Fongching Chau Kevin Fongching Chau Maxim A. Vodyanik Maxim A. Vodyanik Jinlan Jiang Jinlan Jiang Yong Jiang Yong Jiang Characterization of iPSCs derived using the small molecule-aided feeder-free condition. Public Library of Science 2011 ipscs derived molecule-aided feeder-free 2011-03-01 01:25:20 Figure https://plos.figshare.com/articles/figure/_Characterization_of_iPSCs_derived_using_the_small_molecule_aided_feeder_free_condition_/465120 <p>(<b>A</b>) Bright-field image of iPSCs derived from human adult skin fibroblasts (iPS(SK46) clone 2). Scale bar: 100 µm. (<b>B</b>) G-banding chromosome analysis of iPS(SK46) clone 2 (p17). (<b>C</b>) PCR analysis of reprogramming vectors in iPSCs. E: episomal DNA; G: genomic DNA; NF: neonatal foreskin fibroblasts (p5); iPSF7 clone 1 to 3: iPSCs derived from neonatal foreskin fibroblasts (p26); AF: adult skin fibroblasts (p6); iPS(SK46) clone 1 to 3: iPSCs derived from adult skin fibroblasts (p22). piPSC derived from human foreskin fibroblasts (p4) were used as controls. <i>T-OCT4</i>: transgene <i>OCT4</i>; <i>T-SOX2</i>: transgene <i>SOX2</i>; <i>T-NANOG</i>: transgene <i>NANOG; T-LIN28</i>: transgene <i>LIN28</i>; <i>T-c-MYC</i>: transgene <i>c-MYC</i>; <i>T1-KLF4</i>: transgene <i>KLF4 (1)</i>; <i>T2-KLF4</i>: transgene <i>KLF4 (2)</i>; <i>T-SV40LT</i>: transgene <i>SV40LT</i>; <i>OCT4</i>: endogenous <i>OCT4</i>. 32 PCR cycles were used for all primer sets. (<b>D</b>) Quantitative RT-PCR analysis of the endogenous OCT4, NANOG, SOX2 and LIN28 expression in iPSC clones. Data shown are mean ± SEM (n = 3). (<b>E</b>) Bisulfite-sequencing analysis of the methylation status of the <i>OCT4</i> and <i>NANOG</i> promoters in iPSC clones. Open circles indicate unmethylated, and filled circles indicate methylated CpG dinucleotides. (<b>F</b>) Hematoxylin and eosin staining of teratoma sections of iPSC(SK46) clone 2. Teratomas were obtained from all iPSC clones. Left panel: neural tissue (ectoderm); middle panel: cartilage (mesoderm); right panel: gut epithelium (endoderm). Scale bars: 100 µm.</p>