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. 2019 Sep 3;18:1536012119870899. doi: 10.1177/1536012119870899

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© The Author(s) 2019

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).

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Figure 3. — Temozolomide (TMZ) treatment enriches CD133+ glioblastoma multiforme (GBM) cell population with glioma stem cell (GSC) properties. A, Representative fluorescence micrographs (left panels) depict that DBTRG-05MG-CD133-LG post 4-week exposure of TMZ (50 µmol/L) contained an increased CD133+ cell population. Representative flow cytometric analysis shows that TMZ exposure led to a substantial increase in green fluorescent protein (GFP)/CD133+ DBTRG-05MG-CD133-LG cells, from approximately 9.0% to 67%. B, Luciferase assay showed that after 4-week low-dose TMZ exposure, the luciferase activity driven by CD133 promoter in the DBTRG-05MG-CD133-LG was significantly increased as compared to the parental cells. ***P < .001. Comparative colony (C) and neurosphere (D) forming assays. E, Cell viability assay shows that TMZ-treated DBTRG-05MG-CD133-LG became more TMZ resistant (half maximal inhibitory concentration (IC₅₀) value increased from approximately 400-500 µmol/L). E, Western blots depicts the prominently increased expression of stemness markers notch1, β-catenin, and Bruton’s tyrosine kinase (Btk) in the TMZ-treated cells.