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. 2021 Jun 28;49(13):7361–7374. doi: 10.1093/nar/gkab517

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Targeted manipulation of m6A level via dCasRx epitranscriptomic editors on endogenous transcripts affects GSC proliferation. (A) Expression of dCasRx epitranscriptomic editors dCasRx-METTL3 or dCasRx-ALKBH5 in GSC 3565. (B) Representative immunofluorescence images of GSCs transfected with HA-tagged dCasRx epitranscriptomic editors. Scale bars, 40 μm. (C) Methylation at A3488 of endogenous mRNA FOXM1 in GSC 3565 increased by three folds by dCasRx-METTL3 editing. (D) FOXM1 mRNA decreased after dCasRx-METTL3 editing in GSC 3565. (E) Proliferation of GSC 3565 decreased with an increased m6A levels at FOXM1 A3488 mediated by dCasRx-METTL3, compared to NT group and dCasRx-dMETTL3 group. (F) A decrease of methylation at A5553 of endogenous mRNA MYC was mediated by dCasRx-ALKBH5 editing in GSC 3565. (G) mRNA expression level of MYC decreased after dCasRx-ALKBH5 editing in GSC 3565. (H) Proliferation of GSC 3565 decreased with a decreased m6A level at MYC A5553 mediated by dCasRx-ALKBH5, compared to NT group and dCasRx-dALKBH5 group. Data are represented as mean ± SEM. (ANOVA; *, P-value < 0.05; **, P-value < 0.01; ***, P-value < 0.001; ****, P-value < 0.0001, n = 3).