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. 2021 Aug 12;11:16427. doi: 10.1038/s41598-021-95941-4

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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 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/.

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AMPK and AKT regulate Api5 stability. MCF7 cells were treated with (a) 20 μM AMPKi, and (c) 5 μM AKTi for 4 h and 24 h, respectively. Immunoblotting analysis was performed using Api5 specific antibody to check for Api5 expression levels. GAPDH was used as loading control. (b,d) Quantification showing the fold change in Api5 levels after normalisation to GAPDH. (a) p21 and (c) pAkt S473 were used as positive control to confirm AMPK and Akt inhibition, respectively. (e) pAkt S473 activation upon AMPK inhibition. pAkt, Akt and GAPDH blots were cropped and the full blot is provided in Supplementary figure S12. (f) Quantification showing the fold change in pAkt S473 activation after normalisation to GAPDH. (e) E2F1 levels upon AMPK inhibition and (g) Quantification of E2F1 levels after normalising to GAPDH. (h,i) E2F1 protein levels were analysed following Akt inhibition using immunoblotting.