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. 2017 May 15;8:535. doi: 10.3389/fimmu.2017.00535

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Copyright © 2017 Post, Cuapio, Osl, Lehmann, Resch, Davies, Bilban, Schlechta, Eppel, Nathwani, Stoiber, Spanholtz, Casanova and Hofer.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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HOBIT shRNA nearly abrogates formation of CD56⁺ natural killer (NK) cells. Stem cell cultures were transduced and cultured until day 35. The generation of CD56⁺ NK cells was monitored by flow cytometry in samples taken at days 24, 26, 28, 31 and 35. (A) Exemplary flow cytometry dot plots displaying staining for CD56 within the GFP⁺ fractions at days 24 and 35. (B) The formation of CD56⁺ cells within the transduced GFP⁺ and non-transduced GFP⁻ fractions is shown for shHOBIT and shControl cultures. Expansion rates are calculated from measured cell numbers at the respective days and the flow cytometry-derived percentages. Differences in transduction efficiency were corrected for by normalizing the obtained values to the number of GFP⁺ and GFP⁻ cells at day 8, respectively. Results are calculated form four independent experiments, two of them performed in triplicates, using different donor cells and are displayed as mean ± SEM (*p < 0.05, ***p < 0.001).