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. 2018 Oct 31;9:4552. doi: 10.1038/s41467-018-06968-7

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© The Author(s) 2018

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. 7 — PRMT2 methyltransferase activity is required for GBM cell growth and tumorigenesis. a Correlation between H3R8me2a levels and tumor grades. Tumor sections from 21 glioma specimens were IHC-stained with anti-H3R8me2a antibody. Lines within boxes indicate medians of the scores. b Correlation between H3R8me2a levels and Ki67 density in 21 GBM specimens. Representative examples of H3R8me2a and Ki67 immunostainings are shown in different grade of glioma specimens; scale bar, 200 µm (left panel). Stainings of nuclear H3R8me2a and Ki67 were scored and the significance of the correlation was determined by Pearson’s correlation test (right panel). c Cellular levels of PRMT2 and H3R8me2a in U87 and T98G cells grown with or without PRMT2-H112Q expression. In order to induce PRMT2-H112Q expression, cells were treated with or without Doxycycline (Dox 1 mg/l) for 96 h. d Cell growth curves of U87 cells with or without PRMT2-H112Q expression. Significance level was determined using Student’s two-sided t-tests. ***p ≤ 0.001. Error bars, ± SD, n = 3. e Frequency of sphere-initiating cells as measured by limiting dilution analysis in U87 (top panel) or T98G (bottom panel) cells with or without PRMT2-H112Q expression (mean ± SD, n = 3). Frequency and probability estimates were computed using the ELDA software. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001. f The relative mRNA levels of cell cycle associated genes were compared in U87 cells with or without PRMT2-H112Q expression. Significance level was determined using Student’s two-sided t-tests. ***p ≤ 0.01. g Representative luciferase images of three mice per group at 7, 14, and 28 days post tumor implantation. h Survival analysis of mice intracranially implanted with U87 cells with or without PRMT2-H112Q expression. X axis represents days after cells injection. Significance level was determined using log-rank analysis, n = 6 for each treatment group. *p ≤ 0.05. i Moribund mice were killed and dissected tumors were examined by IHC staining in two group specimens with or without PRMT2-H112Q expression. Scale bars, 200 µm