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. 2022 Jul 15;21:146. doi: 10.1186/s12943-022-01607-8

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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. 10 — Knockdown of the m6A reader YTHDF2 promoted the expression of circPOLR2A in cRCC cells. a Pull-down assay validated the binding of YTHDF2 and circPOLR2A. Top, 786O cells; bottom, Caki1 cells. b RIP assay confirmed the binding of YTHDF2 and circPOLR2A. Left, 786O cells; right, Caki1 cells. c RNA FISH and immunofluorescence analysis of the colocalization of YTHDF2 (red) and circPOLR2A (green). d YTHDF2 expression was lower in cRCC tissues than in normal tissues based on TCGA database. e Kaplan-Meier method and log-rank test indicated that YTHDF2 expression was a favorable prognostic factor in the TCGA-KIRC cohort. f MeRIP assay verified the enrichment of circPOLR2A in m6A-precipitated fraction. g-h The expression of circPOLR2A in cRCC cells transfected with siNC, siMETTL3 or siYTHDF2. 10 g, 786O; 10 h, Caki1. i MeRIP assay determined the relative enrichment of circPOLR2A-WT or circPOLR2A-MUT in m6A-precipitated fraction in 786O cells transfected with circPOLR2A-WT or circPOLR2A-MUT plasmids. j Analysis of the relative abundance of circPOLR2A-WT or circPOLR2A-MUT in circPOLR2A-WT-expressing or circPOLR2A-MUT-expressing 786O cells transfected with siNC, siMETTL3 or siYTHDF2. k Analysis on the relative abundance of circPOLR2A in cRCC cells transfected with siNC or siYTHDF2. Blue, cRCC cells were pretreated with DMSO; Red, cRCC cells were pretreated with 10 μM DZNep for 1 h