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

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Figure 5. — Immunocytochemical analyses of hESCs cryopreserved in SCK or DAP for the stem cell markers octamer-binding transcription factor 3/4 (Oct3/4), stage-specific embryonic antigen-4 (SSEA4), and Tra-1-60. Images of hESCs in bright-field, immunofluorescence, and 4′,6-diamidino-2-phenylindole (DAPI) staining were obtained on a microscope at a magnification of 40x. Scale bars: 100 um. hESCs were fixed with 4% PFA, permeabilized, and reacted with primary antibodies against Oct3/4, SSEA4, or Tra-1-60 at 4° C overnight, and further reacted with fluorescently tagged secondary antibodies in the dark. Mounting medium containing DAPI was added, and cells were observed on a fluorescence microscope. (a-c) hESC/SCK/Oct3/4. (d-f) hESC/DAP/Oct3/4. (g-i) hESC/SCK/ SSEA4. (j-l) hESC/DAP/SSEA4. (m-o) hESC/SCK/Tra-1-60. (p-r) hESC/DAP/Tra-1-60.