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. 2020 Jul 28;10:12641. doi: 10.1038/s41598-020-69616-5

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© The Author(s) 2020

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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Immunofluorescence analysis of PD-L1 expression in bone marrow-aspirated plasma cells from patients with multiple myeloma. (A) Formalin-fixed, paraffin-embedded bone marrow aspirate specimens (clot section) from myeloma patients were sectioned at 4–5 µm. The sections were then incubated with antibodies to CD138 (1:100) and PD-L1 (1:100) overnight at 4 °C, followed by incubation with the appropriate secondary antibodies (Alexa Fluor 488, 1:200 and Alexa Fluor 647, 1:200) at room temperature for one hour. Nuclei were counterstained using DAPI, and all images were captured using a confocal laser scanning microscope (CLSM 800, Carl Zeiss Microscopy GmbH). Original magnification × 200. (B) Representative immunofluorescence images for the PD-L1 expression scores and groups based on the predetermined cut-off value of 7.65. (C) The distribution of the PD-L1 expression scores in patients with multiple myeloma. PD-L1 programmed death-ligand 1.