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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. 4 — CircPOLR2A could interact with the PEBP1 protein. a RNA pull-down assay was performed in cRCC cells using a circPOLR2A sense probe (circPOLR2A probe) and an antisense probe (ctrl probe). b Venn diagram of the overlapping proteins (including PEBP1 and SMARCA1) between circPOLR2A-interacting proteins identified by MS and proteins with independent prognostic value in the cRCC cohort from the CPTAC database. c The effect of circPOLR2A on the protein levels of PEBP1 and SMARCA1 was assessed by western blotting. d RNA pull-down assay and western blotting validated the existence of PEBP1 in the precipitates pulled down with the circPOLR2A probe. e RIP assay validated the interaction between PEBP1 and circPOLR2A. Top, western blot analysis for IP efficiency of PEBP1 antibody; bottom, the enrichment of circPOLR2A in the precipitates of PEBP1 antibody relative to input group. f RNA FISH and immunofluorescence analysis indicated the colocalization of PEBP1 (red) and circPOLR2A (green) in 786O cells. g Forest plots showing the prognostic value of PEBP1 protein in the cRCC cohort from the CPTAC database. Top, the results of univariate Cox regression analysis (green dots); bottom, the results of multivariate Cox regression analysis (red dots). h GO enrichment analysis of circPOLR2A-interacting proteins identified via RNA pull-down and MS.