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. 2021 Oct 18;21:541. doi: 10.1186/s12935-021-02257-4

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — DUSP16 is down-regulated in OS and is a downstream target gene of miR-769-5p. A Bioinformatics analysis showed that hsa-miR-769-5p has a total of 14 target genes in miRDB, miRTarBase, and TargetScan; B, C The mRNA expression level of DUSP16 in OS cell lines and hFOB1.19 (B) and clinical samples (C); D, E The protein level of DUSP16 in OS cell lines and hFOB1.19 (D) and clinical samples (E); F Representative immunohistochemical staining of DUSP16 between OS tissues and adjacent tissues; G Negative correlation between miR-769-5p and DUSP16 expression in OS tissues; H Luciferase reporter assay was performed to confirm that miR-769-5p directly bound to the 3′-UTR region of DUSP16. Luciferase activity was analyzed in OS cells co-transfected with miR-769-5p mimics or negative control with pGL3-DUSP16-WT or pGL3- DUSP16-MUT; I, J The expression of DUSP16 in OS cells after alteration of miR-769-5p expression was detected by qRT-PCR (I) and western blot (J); K, L Western blot (K) and immunohistochemical analysis (L) were performed to evaluate DUSP16 expression in vivo; n = 5 mice/group. Data are presented as the means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001