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. 2021 Apr 15;11:8321. doi: 10.1038/s41598-021-87700-2

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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(a) Non-transduced (NT) and LV/hu-IL-12 transduced human sarcoma lines for osteosarcoma (143B), Ewing sarcoma (A673), and rhabdomyosarcoma (RD) were plated in ultralow adherent 96-well plates. After 48 h of growth, media or NK-92mi at a 10:1 ratio were added. Supernatants were assessed by ELISA for IFN-γ. N = 12; 3 independent experiments, displayed mean ± standard deviation. (b) Non-transduced and LV/hu-IL-12 transduced human sarcoma lines were plated. 4 h later conditioned supernatant was collected and 100 µL applied to NK-92mi cells. After an additional 4 h supernatant was collected, and IFN-γ was measured by ELISA. N = 6; 2 independent experiments, displayed mean ± standard deviation. (c) NK-92mi cells were incubated in the presence or absence of anti-IL-12 antibody and 100 µL of conditioned supernatant. After 4 h supernatant was collected, and IFN-γ was measured by ELISA. N = 6; 2 independent experiments, displayed mean ± standard deviation. (d) Primary human NK cells were plated with non-transduced (NT) and LV/hu-IL-12 transduced human Ewing sarcoma (A673) at the noted E:T ratios or conditioned supernatant (100 µL) in the presence or absence of anti-IL-12 antibody. After 6 h supernatant was collected, and IFN-γ was measured by ELISA. N = 3; displayed mean ± standard deviation. * = p < 0.05, ** = p < 0.001, *** = p < 0.0001.