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. 2017 May 1;26(5):773–787. doi: 10.3727/096368916X692654

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Figure 6. — FACS analyses of the expression of Nanog and Tra-1-60 in human embryonic stem cells (hESCs) cryopreserved with SCK or a solution of 2 M DMSO, 1 M acetamide, and 3 M DAP. Freeze-thawed hESCs were cultured on a layer of feeder cells for 5 days, disaggregated by TrypLE Select, fixed with 4% PFA, and permeabilized. Cells were then reacted with primary antibodies against Nanog and Tra-1-60 for 1 h and further reacted with fluorescently tagged secondary antibodies in the dark. The control was hESCs without primary antibody treatment. hESCs were analyzed using the ABI FACSCalibur. Data were analyzed by Quest software and presented as the control (open histogram), and samples were treated with primary antibodies (closed histogram). (a) hESC/SCK/ Nanog. (b) hESC/DAP/Nanog. (c) hESC/SCK/Tra-1-60. (d) hESC/DAP/Tra-1-60.