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. 2022 Dec 6;41:334. doi: 10.1186/s13046-022-02538-4

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 8 — PFKL positively promotes YTHDF3 expression at protein level and PFKL interacts with YTHDF3 via EFTUD2 at the protein level. a PFKL and YTHDF3 protein expression by transiently transfecting small interfering RNA targeting PFKL mRNA into Sk-Hep-1, HepG2 and SNU449 cells for 12 hours. b. PFKL and YTHDF3 protein expression by transiently transfecting PFKL plasmids into Sk-Hep-1 and HepG2 cells for 12 hours. c-f. CCK8 proliferation assays, colony formation assays, transwell migration assays and transwell invasion assays after transiently transfecting small interfering RNA targeting PFKL into overexpression-YTHDF3 Sk-Hep-1, HepG2, SNU449 cells for 12 hours. The Image J software was used to quantified cells’ migration and invasion ability. Scale bar 200 μm. g. Endogenous co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL with Huh7 and SNU449 cells. h Exogenous co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL with YTHDF3-stably-overexpressed Sk-Hep-1 and HepG2 cells. i Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL after co-transfecting YTHDF3 and PFKL plasmids (10 μg respectively) into 293 T cells for 48 h. j The processes of PFKL pre-mRNA splicing and YTHDF3 m⁶A modification to PFKL mRNA. k-m Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL after transfecting EFTUD2 plasmids (10 μg) into Sk-Hep-1, Huh7 and SNU449 cells for 48 h. n Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and EFTUD2 after co-transfecting YTHDF3 and EFTUD2 plasmids (10 μg respectively) into 293 T cells for 48 h. o Co-immunoprecipitation assays were performed to investigate the interaction between PFKL and EFTUD2 after co-transfecting PFKL and EFTUD2 plasmids (10 μg respectively) into 293 T cells for 48 h

p Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL after co-transfecting YTHDF3, PFKL and EFTUD2 plasmids (10 μg respectively) into 293 T cells for 48 h. Data are presented as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001).