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. 2021 Oct 6;53(10):1547–1558. doi: 10.1038/s12276-021-00674-z

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© The Author(s) 2021

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 Left, circSPG21 and probe diagram. Right, RNA-fluorescence in situ hybridization (FISH) showed that circSPG21 was mainly present in the cytoplasm (scale bar, 100 μm). The circular RNA probe was labeled with Cy-3. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). b Nucleus pulposus cells were treated with 5 μg/mL actinomycin D, and the expression levels of circSPG21 and SPG21 were detected by RT-qPCR, *p < 0.05. Data are presented as the mean ± S.D., and the p values were determined by a two-tailed unpaired Student’s t-test. c Exons 2–3 of SPG21 form circSPG21. Sanger sequencing confirmed the presence of circSPG21. d Left: circSPG21 was amplified by divergent and convergent primers in cDNA and genomic DNA and separated by horizontal electrophoresis. β-actin served as a negative control. Right: a diagram of the divergent and convergent primers.