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. 2021 Jun 6;20:85. doi: 10.1186/s12943-021-01366-y

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Rigosertib promotes T cell and NK cell responses but attenuates tumor-associated M2 macrophages in the tumor microenvironment. YUMM3.3 tumors were collected 17 days post-treatment. a, h Live CD45⁺ leukocytes were concatenated after downsampling to 20,000 events for subsequent high-dimensional data analysis to normalize the contribution among samples under different treatments. Samples were then analyzed in parallel by t-SNE and manually gated leukocyte populations were overlaid onto the total t-SNE map using FlowJo 10.5.3. b-e, g, i Flow cytometric and f IHC analysis of YUMM3.3 tumors at day 17 post-treatment. j Flow cytometric tSNE analysis of PBMC samples from YUMM3.3 tumor-bearing mice treated with RGS before and after CD4 and/or CD8 antibody depletion treatments. (K) Tumor weight (day 16 post RGS treatment) of YUMM3.3 tumors in C57BL/6 mice with and without CD4 and/or CD8 depletion. a-i pooled data obtained from at least two different experiments (n = 5 ~ 10) are shown. j, k were replicates (n = 10 per group)