Journal: Cell Death & Disease

Article Title: Short-term retinoic acid treatment sustains pluripotency and suppresses differentiation of human induced pluripotent stem cells

doi: 10.1038/s41419-017-0028-1

Figure Lengend Snippet: List of primers used for qRT-PCR

Article Snippet: Cells were further incubated for 3 h in blocking solution with the following primary antibodies: human Nanog (1:1000; rabbit polyclonal, PA1-097, Thermo Fisher Scientific), human Oct4 (1:400 mouse monoclonal, 60093, STEMCELL Technologies), human β-catenin (1:200 mouse monoclonal, sc-7963, Santacruz), human TRA-1-60 (1:100 mouse monoclonal, 41–1000, Thermo Fisher Scientific), human Brachyury (1:20 goat polyclonal, AF2085, R&D systems), human Sox17 (1:20 goat polyclonal, AF192, R&D systems) and human Nestin (1:1000 mouse monoclonal, 60091, STEMCELL Technologies).

Techniques: Sequencing

Journal: Cell Death & Disease

Article Title: Short-term retinoic acid treatment sustains pluripotency and suppresses differentiation of human induced pluripotent stem cells

doi: 10.1038/s41419-017-0028-1

Figure Lengend Snippet: a Representative image of EBs formation capability of RA-treated and untreated hiPSCs-F and -TL. Scale bar, 500 μm. b Diagram indicating the average diameter of the EBs, 100 μm. c The efficiency of EBs formation with diameter range of 100–300 µm is higher in hiPSCs-RA 8 compared to untreated cells, while the percentage of EBs with diameter range of 50–100 μm is comparable between hiPSCs-RA 8 and hiPSCs. d The potential of hiPSCs-RA 8 to differentiate into cell of all three germ layers by expression analysis of HAND1 , NESTIN , and SOX17 , the gene expression levels in EBs hiPSCs and EBs hiPSCs-RA 8 were relative to hiPSCs. qRT-PCR data are represented as the mean ± SEM from three independent experiments. A statistical comparison was made between EBs hiPSCs-RA 8 and EBs hiPSCs-RA by Student’s t -test (** p < 0.01, *** p < 0.001). e Immunostaining of whole EBs for Brachyury (upper panel), Sox17 (middle panel), and Nestin (bottom panel) demonstrating the differentiation potential of hiPSCs-RA 8. Scale bar, 50 μm

Article Snippet: Cells were further incubated for 3 h in blocking solution with the following primary antibodies: human Nanog (1:1000; rabbit polyclonal, PA1-097, Thermo Fisher Scientific), human Oct4 (1:400 mouse monoclonal, 60093, STEMCELL Technologies), human β-catenin (1:200 mouse monoclonal, sc-7963, Santacruz), human TRA-1-60 (1:100 mouse monoclonal, 41–1000, Thermo Fisher Scientific), human Brachyury (1:20 goat polyclonal, AF2085, R&D systems), human Sox17 (1:20 goat polyclonal, AF192, R&D systems) and human Nestin (1:1000 mouse monoclonal, 60091, STEMCELL Technologies).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Comparison, Immunostaining