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. 2022 Oct 7;58(10):1411. doi: 10.3390/medicina58101411

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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The effect of co-culturing with C1R, C1R-B*2704 C67S, or C1R-B*2704 cells on the cytokine expression of NK-92MI cells. After co-culturing with C1R, C1R-B*2704, or C1R-B*2704 C67S, NK-92MI cells were isolated by using the EasySep™ Human NK Cell Isolation Kit. The total RNAs of treated NK-92MI cells were isolated by using the QIAamp RNA Blood Mini kit (QIAGEN, GmbH, Germany). The results were averaged from four independent experiments. The levels of IFN-γ (A), TNF-α (B), or IL-6 (C) mRNA of NK cells were analyzed through quantitative RT-PCR. Real-time RT-PCR values for each cytokine were normalized to those of 18S rRNA. Relative expression levels of mRNA were calculated by the following equation: (40−threshold cycle [Ct], adjusted by the expression of 18S rRNA).