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. 2020 Jun 15;11:3015. doi: 10.1038/s41467-020-16827-z

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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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Fig. 2 — a Immunoblot analysis of WISP1 expression in GSCs transduced with non-targeting shRNA (shNT) or WISP1 shRNA (shWISP1) through lentiviral infection. b Cell viability assay of GSCs transduced with shNT or shWISP1. n = 6 (T4121 GSCs) or 5 (T387 GSCs) biological independent samples. Data are shown as means ± s.d. ****p <0.0001, two way ANOVA analysis followed by Tukey’s multiple test. c Representative tumorsphere images of GSCs transduced with shNT or shWISP1. Scale Bar: 200 μM. d Quantification of tumorspheres formed by GSCs transduced with shNT or shWISP1. n = 6 (T4121 GSCs) or 5 (T387 GSCs) biological independent cell cultures. Data are shown as means ± s.d. ****p <0.0001, two-tailed unpaired t-test. e In vitro extreme limiting dilution analysis of the tumorsphere formations of GSCs expressing shNT or shWISP1. n = 30 biological independent cell cultures. ****p <0.0001 by ELDA analysis. f Cell viability assay of GSCs transduced with shNT or shWISP1 and cultured with different dose of recombinant human WISP1 (rWISP1) protein for 4 days. n = 6 biological independent samples. Data are represented as means ± s.d. ****p <0.0001, two-tailed unpaired t-test. g Immunoblot analysis of WISP1 expression in T4121 GSCs after transduction with WISP1 overexpression or vector control. h Cell viability assay of T4121 GSCs transduced with WISP1 overexpression or vector control. n = 6 biological independent samples. Data are represented as means ± s.d. **p = 0.0014, ****p <0.0001, two-tailed unpaired t-test. i Tumorsphere formation of T4121 GSCs transduced with WISP1 overexpression or vector control. n = 6 biological independent cell cultures. Data are shown as means ± s.d. ****p <0.0001, two-tailed unpaired t-test. Source data are provided as a Source data file.