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. 2022 Jun 11;8:285. doi: 10.1038/s41420-022-01066-6

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© The Author(s) 2022

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. 5 — A The network of target genes of miR-27a and miR-27b, the red circle represents PPARγ. B Schematic illustration showed binding site sequences between PPARγ and miR-27a/27b. C RIP assay showed PPARγ mRNA was effectively pulled down by AGO2 with a much greater enrichment in UM1 cells. D RNA pull-down assay exhibited for the direct bonding of PPARγ and miR-27a/27b in UM1 cells. E Dual-luciferase assays were conducted using PPARγ wild type (WT) and mutant (MUT) 3′UTR reporter constructs to show that the human PPARγ 3′UTR region contains miR-27a and miR-27b binding sites. (Data were normalized to that with the control probe, showing the mean ± SD of three experiments). F Western Blot assay showed miR-27a and miR-27b mimic could significantly reduce expression of PPARγ in UM1 and HN31 cells at the level of proteins. G, H RNA FISH showed colocalization of PPARγ and miR-27a/27b in UM1 and HN31 cells (Scale bar, 10 μm). I, J Pearson correlation analysis indicated miR-27a and miR-27b expression was negatively correlated with PPARγ expression in patients with HNSCC (n = 15 and 15, respectively). Data were shown as the mean value ± SD of three independent experiments. The asterisks indicate significant differences (Student’s t-tests, *P < 0.05, **P < 0.01, ***P < 0.001).