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. 2020 May 22;11:2578. doi: 10.1038/s41467-020-16306-5

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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. 6 — a Schematic representation of positions of m⁶A site within PDK4 mRNA and the regions targeted by three gRNAs, respectively. b The threshold cycle (Ct) of qPCR showing SELECT results for detecting m⁶A site in the potential m⁶A site of PDK4 5′UTR in Mettl3^Mut/- HeLa, sh-Mettl3 Huh7 and their corresponding control cells. c The threshold cycle (Ct) of qPCR showing SELECT results for detecting m⁶A site in 5′UTR of PDK4 in HeLa cells transfected with dCas13b-ALKBH5 combined with gRNA control or gRNA1/2/3, respectively, for 24 h. d, e The mRNA (d) or protein (e) expression of PDK4 in HeLa cells transfected with dCas13b-ALKBH5 combined with gRNA control or gRNA1/2/3, respectively, for 24 h. f RIP-qPCR analysis of PDK4 mRNA in HeLa cells transfected with dCas13b-ALKBH5 combined with gRNA control (dC-A5) or gRNA for PDK4 (dC-A5 + gRNA) for 24 h by use of antibodies against YTHDF1 and IGF2BP3, respectively. g HeLa cells were transfected with gRNA control, gRNA1 for PDK4, and dCas13b-ALKBH5 for 24 h and then further treated with Act-D for the indicated times. The mRNA of PDK4 was checked by qRT-PCR. h–j The glucose consumption (h), lactate production (i), and ATP levels (j) in HeLa cells transfected with gRNA control, gRNA1 for PDK4, and dCas13b-ALKBH5 or dCas13b-ALKBH5-Mut for 24 h. Data are presented as means ± SD from three independent experiments. **p < 0.01, NS, no significant, by two-tailed unpaired Student’s t test for (b) (p = 0.001 and p = 0.002, respectively), (h–j) one-way ANOVA for (c) (p < 0.001), (d) (p < 0.001), and (e) (p < 0.001), and two-way ANOVA for (f) and (j) (p < 0.001).