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. 2022 Oct 20;41:307. doi: 10.1186/s13046-022-02518-8

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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. 3 — CircLRFN5 inhibits GSCs viability and proliferation in a ferroptosis-dependent manner in vitro. a Cell viability assays showed whether VAD (20 μM), Nec (20 μM), 3-MA (60 μM) and Fer-1 (10 μM) could rescue the growth inhibition of GSC51 induced by circLRFN5 overexpression after 48 h. b, c Representative images of BODIPY (581/591) C11 staining in different groups of GSCs and the relative fluorescence intensity of O-BODIPY was quantified by image J. Scale bar = 50 μm. d, e GSH (d) and MDA (e) levels were detected in circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. f, g MTS assays showed the cell viabilities of circLRFN5 overexpressed GSC51 (f) and GSC58 (g), followed by Fer-1 treatment. h, i Representive EdU assays showed the proliferation of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. Scale bar = 50 μm. j Representive neurospheres formation assays showed the relative sizes of neurospheres of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. Scale bar = 20 μm. k ELDA assays showed the neurospheres formation abilities of circLRFN5 overexpressed GSC51 (left) and GSC58 (right), followed by Fer-1 treatment. l Western blotting detects the stemness markers of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. m Cell cycle assays showed the cell cycle distributions of circLRFN5 overexpressed GSC51 and GSC58, followed by Fer-1 treatment. All data are expressed as the mean ± SD (three independent experiments). *p < 0.05; **p < 0.01; ***p < 0.001