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. 2021 Jun 18;11:12893. doi: 10.1038/s41598-021-92223-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 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 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/.

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Immunostaining and Immunohistochemistry of patient-derived AT/RT cells and tissues. (a,b) Two resected patient AT/RT demonstrate strong immunostaining with HML-2 Envelope (Env) monoclonal antibody (brown). (c) Cerebrum from a normal brain is negative when stained with the same antibody. (d–l) AT/RT cell lines express markers of multiple stages of differentiation; representative images are shown. (d) CHLA 02, a patient derived AT/RT cell line, expresses Pax6 (red), a marker for neuroectoderm, merged with immunostaining for HML-2 Env (green). (e): This cell line also expresses Nestin (green), a marker of neural stem cells. (f) The same cell line expresses Oct4 (red), a pluripotency marker. (g,h) CHLA 04 cells, another AT/RT line, express Oct4 (green) and Nestin (red). (i) CHLA 05 cells also express Oct4 (green) and Nestin (red). (j) The same cell line also expresses βIII tubulin (red), a marker for neurons and HML-2 envelope (green). (k) CHLA 06 cells also express Nestin (red) and Oct4 (green). (l) The same cell line also expresses βIII tubulin (red) and HML-2 Env (green). Nuclei are stained with DAPI (blue). For IHC images, slides were scanned with a OptraScan Digital Pathology Scanner and processed in Adobe Photoshop for incorporation into the figure. For immunofluorescent images, an EVOS fluorescence microscope (AMG) was used. Images were acquired with the native software installed on the EVOS microscope and processed in Microsoft PowerPoint (https://www.thermofisher.com/us/en/home/technical-resources/software-downloads/evos-fl-cell-imaging-system.html).