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. 2021 Sep 8;20:114. doi: 10.1186/s12943-021-01412-9

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© The Author(s) 2021

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 — CircMYH9 promoted SG metabolism though p53-mediated upregulation of PHGDH. A Heatmap of the differentially expressed genes after circMYH9 knockdown in HCT116 cells. B Bubble chart showing the GO analysis of the differentially expressed genes after circMYH9 knockdown. C-D GSEA showed that genes differentially expressed in response to circMYH9 knockdown were enriched in gene sets of SG metabolism and p53 pathway. E–F SG biosynthetic pathway genes and the neutral amino acid transporter SLC1A4 were detected by qRT-PCR in circMYH9-depleted or circMYH9-overexpressing CRC cells. G The expression of p53, PHGDH, PSPH and SLC1A4 were detected by IB in circMYH9-depleted or circMYH9-overexpressing CRC cells. H-I HCT116 cells expressing a control or circMYH9-shRNA and LoVo cells expressing vector or circMYH9 plasmid were cultured in complete medium with uniformly labelled [U-13C] glucose for 24 h. LC–MS was used to detect the relative intracellular levels of 13C-labelled serine or glycine. The mean value of the peak area ± SD represents the serine and glycine peaks on the MS chromatogram from 5 times of independent experiments(*, P < 0.05; **, P < 0.01). J IB analysis of PHGDH, p53 and MDM2 expression in the LoVo cells treated with circMYH9 overexpression plasmid or/and Nutlin-3. K IB analysis of PHGDH and p53 expression in the circMYH9-depleted HCT116 cells treated with shRNA ctrl or p53 shRNA. L IB analysis of PHGDH and p53 expression in the circMYH9-overexpressing LoVo cells treated with shRNA ctrl or PHGDH shRNA. M IHC staining of p53, PHGDH and PSPH in xenograft tumours transfected with control or circMYH9-depleted HCT116 cells. N IHC staining of p53, PHGDH and PSPH in xenograft tumours transfected with vector or circMYH9-overexpressing LoVo cells. Data (except H and I) are shown as the mean ± SD of three independent experiments (*, P < 0.05; **, P < 0.01)