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. 2020 Oct 28;11(10):925. doi: 10.1038/s41419-020-03126-0

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© The Author(s) 2020

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. 3 — A SA-β-gal staining (100×) showed that remarkable senescence in cells was elicited by the CDK4/6 inhibitor LY2835219 (Cal27: 1.25 μM; HSC6: 0.3 μM; HSC3: 0.1 μM) but not by metformin (Cal27: 10 mM; HSC6: 1 mM; HSC3: 10 mM). Compared with LY2835219 monotherapy, the combination including metformin had no significant impact on the proportion of senescent cells. Bar: 400 μm. B Antibody array results indicated that the levels of a series of tumor-promoting SASP factors (such as IL6, IL8, MCP1, and GRO) were upregulated by LY2835219, while the combination including metformin inhibited this upregulation. C qRT-PCR results confirmed the modulation of the SASP by metformin. D Western blot results showed that the mTOR and stat3 pathways were activated by LY2835219. E Metformin treatment inhibited the mTOR and stat3 pathways. F The combination treatment inhibited the stat3 and mTOR pathways, which were activated by LY2835219 monotherapy. G qRT-PCR results confirmed that both INK-128 (a selective mTOR inhibitor) and S3I-201 (a selective stat3 inhibitor) could suppress the upregulation of the expression of some of the SASP factors induced by LY2835219. *P < 0.05 when compared with the control group; ^#P < 0.05 when compared with the combined group; ns indicates no significant difference; one-way ANOVA. LY: LY2835219; Met: metformin.