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. 2021 May 10;40:162. doi: 10.1186/s13046-021-01936-4

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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. 5 — Timeless activates Wnt/β-catenin signaling. Western blotting was performed to detect total (a) and nuclear β-catenin (b) in Timeless knockdown and overexpressing CRC cells. (c) The laser scanning confocal microscope was performed to detect the subcellular location of β-catenin. Original magnification, × 400 (d) Expression levels of Axin2, CCND1, LEF1 and CTNNB1 were assayed by qRT-PCR in the indicated cells. An immunoprecipitation (IP) assay was performed to detect Flag-tagged β-catenin (e) or (f) Flag-tagged Timeless that interacted with Timeless or β-catenin in HCT116 cells. Error bars indicate the means ± SD. *P < 0.05, **P < 0.01