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. 2022 Feb 23;7:52. doi: 10.1038/s41392-022-00872-9

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© The Author(s) 2022

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. 3 — Single-Cell RNA-Seq Reveals activated microglia in metastatic lesions. a Schematic representation of the in vivo selection process. Parent A549 cells were inoculated into the left cardiac ventricle of nude mice. Tumor cells were isolated from brain lesions and reinoculated after expansion in culture. Cells isolated from the second round of metastases were expanded in culture and reinoculated to get the third-generation cells. b Histogram showed incidence of brain metastases in different cell lines (parental: 10.0%; A549-F1: 28.6%; A549-F2: 50.0%; A549-F3: 63.6%). c Schematic overview of the experimental design for the single-cell RNA-seq analyses was generated from the online tool (BioRender: https://biorender.com/). d UMAP analyzed cells reveal the existence of 20 clusters within the three experimental groups (untreated mouse brain, brain metastases from A549 and A549-F3 cells). e UMAP plots indicated distribution of cell clusters in different groups. f The numbers of different cell types in each group. g The numbers and percentages of anti-inflammatory and pro-inflammatory microglia in different groups. h The top 20 signal pathways with significant difference showed in KEGG