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. 2023 Sep 12;12(1):2240670. doi: 10.1080/2162402X.2023.2240670

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© 2023 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 3. — hPSC-NK Cell Characterization.

a. PhC image showing the morphologic appearance of hPSC-NKs. b. PhC image showing the morphologic appearance of PB-NKs. c. PhC image showing the morphologic appearance of activated hPSC-NKs. d. Immunofluorescent image demonstrating granules containing Granzyme B (green) within the cytoplasm of hPSC-NKs. DAPI (blue) is used to visualize cell nuclei. e. Flow cytometric analysis showing CD56, CD16, CD94, NKp46, and NKG2A hPSC-NK marker expression. f. Flow cytometric analysis showing high expression of perforin in hPSC-NKs. g. Flow cytometric analysis demonstrating hPSC-NK production of cytokines, TNFα, and IFNγ. Cells were stimulated with PMA/ionomycin in the presence of Brefeldin A (overnight), fixed, permeabilized, and stained for TNFα or IFNγ. h. MetaCore analysis demonstrating the top 50 statistically significant functions among PB-NKs and hPSC-NKs. i. A category network plot showing statistically significant genes (RNA-Seq) that play an important role in NK cell anti-tumor response. j. Flow cytometric analysis showing that hPSC-NKs exhibit high cytotoxicity against K562 cells; PI = propidium iodide. k. PhC image (left) of Jurkat cells co-incubated with hPSC-NKs for 2 hrs. PI staining (right) showing that the majority of Jurkat cells are non-viable.