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. 2021 Oct 23;10(10):70. doi: 10.1038/s41389-021-00362-7

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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 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. 5 — A, B ACTL6A-knockdown increases p53 and p21^Cip1 mRNA levels and p21^Cip1 protein level, but p53 protein is not detected in ACTL6A competent or knockdown Meso-1 cells. C Chromatin IP assay shows ACTL6A binding to the distal p53 binding sites and the proximal promoter Sp1 binding sites in Meso-1 cell extracts, but no binding of p53 is detected. D p53 knockdown does not alter Meso-1 cell spheroid number or invasion. E, F p53 protein is absent in ACTL6A knockout cells even though p53 mRNA levels are increased. G ACTL6A knockdown increases p53 in NCI-Meso-17 cells. H ACTL6A binds to the p21Cip1 promoter Sp1 and p53 sites. p53 binds to the p53 binding sites (p21-p53 sites 1 and 2) but not the Sp1 binding sites (p21-proximal). I, J ACTL6A knockdown reduces spheroid formation and invasion, p53 knockdown increases spheroid formation and invasion, and combination treatment with ACTL6A- and p53-siRNA does not alter ACTL6A suppression of spheroid formation and invasion. This suggests that ACTL6A suppresses the ability of p53 to activate p21^Cip1 promoter transcription. K ACTL6A knockdown does not increase p53 mRNA level. Single asterisks indicate a significant decrease (n = 3, p ≤ 0.001) and double asterisks indicate a significant increase (n = 3, p ≤ 0.001). All statistical comparisons are to the control group. Single asterisks indicate a significant decrease (n = 3, p ≤ 0.001) and double asterisks indicate a significant increase (n = 3, p ≤ 0.001).