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. 2022 Jul 25;54(7):1024–1037. doi: 10.1038/s12276-022-00806-z

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© The Author(s) 2022

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. 2 — a The complementary sequences of intron 2 of ROCK1 were blasted with reverse sequences of intron 4 of ROCK1. Very highly similar sequences (83% identity over 294 nt) were found, as illustrated by using BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi). b Four vectors (vector #1 containing FIS2 and FIS4; vector #2 with FIS2 deletion; vector #3 with FIS4 deletion; vector #4 with FIS2 and FIS4 deletion) were transfected into U2OS and HOS cells, and the expression level of circROCK1-E3/E4 was checked by an RT–qPCR assay. ^****p < 0.0001. c QKI was knocked down as determined by a western blot assay. d Expression of QKI mRNA, circROCK1-E3/E4 and ROCK1 mRNA after transfection of QKI siRNAs was analyzed by an RT–qPCR assay. ^n.sp > 0.05 and ^**p < 0.01. e By using IgG as a control, partial segments of intron 2 and intron 4 of ROCK1 rather than β-actin were abundantly enriched in QKI, as measured by an RIP assay. ^n.sp > 0.05 and ^****p < 0.0001. All data are presented as the mean ± SD from three independent experiments.