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. 2017 Aug 11;7:7958. doi: 10.1038/s41598-017-08483-z

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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PD-L1/NKG2D ligand expressions in cisplatin-resistant cells vs. parental cells. (A) qPCR (upper panel) and Western blot (lower panel) analyses detecting expressions of PD-L1 in A549P/A549CisR and H157P/H157CisR cells. (B) Flow cytometric analyses of PD-L1 in A549P/A549CisR and H157P/H157CisR cells. Relative fluorescence stained with APC-conjugated anti-PD-L1 Ab was shown. (C) Western blot analysis of PD-L1 in A549P/A549CisR and H157P/H157CisR cells after co-culture with NK cells. Upper panel shows the PD-L1 levels in A549P/A549CisR and H157P/H157CisR cells after co-culture with NK cells for 4 hours. Lower panel shows the PD-L1 levels in A549P/A549CisR and H157P/H157CisR cells after incubation (4 hours) with recombinant human IFNγ (rhIFNγ). (D) IHC staining of PD-L1 in tumor tissues of H157P and H157CisR-derived xenografts. PD-L1 levels in tumor tissues, derived from H157P cell and H157CisR cell-derived xenografts were examined. Error bars and significance values were obtained by counting positively stained cells in one randomly chosen phase of slides of 3 different stains. Magnification, 20X. (E) qPCR analyses of NKG2D ligands in A549P/A549CisR and H157P/H157CisR cells. **p < 0.01, ***p < 0.001.