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. 2017 Jun 27;7:4281. doi: 10.1038/s41598-017-04368-3

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© The Author(s) 2017

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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Crosstalk between USP3 and SMAD4 was 3′UTR and miRNA dependent. (a,b) USP3 3′UTR or SMAD4 3′UTR mutually modulated protein expression of the other when endogenous USP3 or SMAD4 expression was first reduced by siRNA transfection. (c,d) Deletion of USP3 or SMAD4 attenuated luciferase activity of the PGL3-USP3 3′UTR or PGL3-SMAD4 3′UTR reporters; (e,f) overexpression of USP3 3′UTR or SMAD4 3′UTR increased the luciferase activity of the corresponding reporter plasmids. (g,h) USP3 and SMAD4 expression was elevated when Dicer depletion was combined with USP3 or SMAD4 knockdown, compared with treatment of si-USP3 or si-SMAD4 alone. All values are represented as the mean ± SD (n = 3). Western blot images are cropped; full-length blots are included in Supplementary Figure S11 to S14. Quantitative protein expression data were normalized to β-actin. *P < 0.05.