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. 2021 Nov 19;6:397. doi: 10.1038/s41392-021-00784-0

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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 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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Fig. 1 — Overexpression of GOLM1 is associated with the immunosuppressive microenvironment in HCC and immune-escape of tumor cells. a Differences in the immunophenotypes of infiltrating immune cells detected by flow cytometry in HCC tissues with high or low GOLM1 expression. t-Distributed Stochastic Neighbor Embedding (tSNE) map derived from flow cytometric analysis indicated the populations of the infiltrating immune cells, including CD3⁺ T cells (CD45⁺CD3⁺), natural killer (NK) cells (CD45⁺CD3⁻CD56⁺), natural killer T (NKT) cells (CD45⁺CD3⁺CD56⁺), monocytes (CD45⁺CD14⁺), macrophages (CD45⁺CD14⁺CD68⁺), and myeloid-derived suppressor cells (MDSC) (CD45⁺CD11b⁺CD33⁺). Percentages of immune cell populations within CD45⁺ cells are shown on the bottom panel (n = 32). b PD-L1 expression profiles detected by flow cytometry in HCC tissues with high or low GOLM1 expression. tSNE map derived from flow cytometric analysis indicated the intensity of PD-L1 expression in CD45⁺ cells. Mean fluorescence intensity (MFI) of PD-L1 staining in tumor and stromal cells (CD45⁻), monocytes (CD45⁺CD14⁺), macrophages (CD45⁺CD14⁺CD68⁺) is shown on the bottom panel (n = 32). c Representative tSNE map derived from flow cytometric analysis of CD8⁺ T cells in HCC tissues with high or low GOLM1 expression. d The functional state of CD8⁺ T cells was analyzed by flow cytometric quantification of GZMB⁺, IFN-γ⁺, PD-1⁺, TIM3⁺, Ki67⁺, and active-caspase3⁺ cells in HCC tissues with high or low GOLM1 expression (n = 32). e The differences of CD68 (macrophages), CD8 (CD8⁺ T cells), and PD-L1 levels detected by immunohistochemical staining (IHC) in human HCC tissues with high or low GOLM1. Representative IHC images for GOLM1, CD68, PD-L1, and CD8 expression in HCC tissues with high or low GOLM1 expression (n = 60) are shown on the top panel. The statistical analysis of IHC score is shown on the bottom panel. f The levels of GOLM1 and PD-L1 were detected by Western blot in HCC tissues and normalized against those of GAPDH using a computerized image system (Image-Pro Plus 6.0). Intensity higher than the median level was defined as high expression. The statistical analysis of quantification is shown on the bottom panel (n = 44). *P < 0.05, **P < 0.01, ***P < 0.001, ns: no significant