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. 2020 Apr 16;10:6499. doi: 10.1038/s41598-020-63403-y

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© The Author(s) 2020

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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Correlation of CREB1 and newly identified CREB1-specific miRNAs. (A) In silico prediction of miRNA promoter and identification of CREB1 responsive elements (CRE sites, red asterisk) within these sequences. Proximal promoter reflects 1,000 bp. (B) Western blot-based detection of CREB1 and densitometric quantification after transfection of esiRNAs specific to CREB1 in MZ2862RC cell line. The number in parentheses indicates a technical replicate. Same blot was re-probed for β-Actin which served as loading control. Uncropped blot is shown in Supplementary Figure 6. (C) Effect of esiRNA mediated CREB1 knock-down on the expression level of indicated miRNAs. miR-9-5p was used as positive control (n = 3, student t-test). (D) Expression of the indicated miRNAs was determined in 18 CREB1 negative and 18 CREB1 high ccRCC lesions by qRT-PCR and results are visualized as Box-Whisker-Plots (Holm-Sidak method, miR-22: p = 0.269; *p < 0.05).