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. 2021 Dec 14;41(7):1063–1069. doi: 10.1038/s41388-021-02147-z

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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. 1 — A Genome browser snapshot of the DDIT4 gene locus. Upper black tracks display publicly available p53 binding signals from Nutlin-3a-treated U2OS [40] and HCT116 [39] cells. Gray and orange tracks display RFX7 binding signals from respective dimethyl sulfoxide (DMSO) and Nutlin-3a-treated U2OS, HCT116, and RPE-1 cells [31]. B ChIP-qPCR of p53 and RFX7 binding to GAPDH (negative control), MDM2 (p53 positive control), and DDIT4 from U2OS cells treated with 10 µM Nutlin-3a or DMSO solvent control. Mean and standard deviation is displayed. Statistical significance obtained through a two-sided unpaired t-test, n = 3 technical replicates. C RT-qPCR data of DDIT4 and RFX7 in 10 µM Nutlin-3a and DMSO control-treated U2OS cells transfected with two different control siRNAs (siCtrl) and two different siRNAs against RFX7. Normalized to siControl#1 DMSO and ACTR10 control gene. Mean and standard deviation is displayed. Statistical significance obtained through a two-sided unpaired t-test, n = 9 replicates (three biological with three technical each). D Western blot analysis of RFX7, DDIT4, p53, and actin (loading control) levels in U2OS, HCT116, and RPE-1 cells transfected with siControl, siRFX7, or siTP53 and treated with DMSO solvent control, 10 µM Nutlin-3a (N3A), 5 nM Actinomycin D (AD), and 1 µM Doxorubicin (Dox).