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. 2018 Oct 10;50(10):132. doi: 10.1038/s12276-018-0160-8

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© The Author(s) 2018

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. 2 — a Synergistic effects of p53 and MSK1 in endogenous p21 transcription. p53-null H1299 cells were transfected with a combination of plasmids encoding transcription activators and MSK1. Cells for ectopic RAR/RXR expression were treated with 9-cis-retinoic acid. b General coactivator functions of MSK1 in p53 target gene expression. H1299 cells were transfected with p53 and MSK1 expression plasmids as indicated. c Requirement of endogenous MSK1 for p21 expression. MCF-7 cells pretreated with nonspecific (siCtrl) and MSK1 (siMSK1) siRNA, as indicated, were treated with 30 μM cisplatin. Cell extracts were subjected to immunoblotting analysis with the indicated antibodies. d Activation of the MSK1 pathway enhances endogenous p21 transcription. H1299 cells were transfected with vectors encoding a combination of p53, MSK1, and a constitutively active MKK6 mutant (MKK6ca) as indicated. a, b, and d Endogenous mRNA levels were measured via one step reverse-transcriptase (RT)-coupled qPCR and normalized relative to β-glucuronidase expression. Error bars represent the mean SD of triplicate samples. The significance of the differences in p21 expression (d) was evaluated using Student’s t-test. Comparable levels of ectopic protein expression were observed in all the examined samples (data not shown)