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. 2022 Oct 9;12:171. doi: 10.1186/s13578-022-00908-1

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© The Author(s) 2022

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. 5 — YAP1 inhibits miR-124-3p and miR-188-5p expression by recruiting YY1 to the promoters. A, B RT-qPCR analysis of the mRNA levels of DCLK1, Vimentin and concurrent expression levels of miR-124-3p and miR-188-5p in HCT116 and SW620 cells transfected with YY1 siRNAs. C Western blot analysis of the expression of YY1, DCLK1 or Vimentin in HCT116 and SW620 cells transfected with two different YY1 siRNAs. β-actin was used as an internal control. D Western blot analysis of YY1, DCLK1 or Vimentin in HCT116 and SW620 cells transfected with YY1 siRNA and then rescued with HA-YY1 plasmid. E RT-qPCR analysis of miR-124-3p and miR-188-5p levels in HCT116 cells transfected with YY1 siRNA and HA-YY1 plasmid compared with siNC and vector control respectively. Bars are means ± SD; *P < 0.05, **P < 0.01, ***P < 0.001, n = 3. F Analysis of the interaction of endogenous YAP1 with YY1 and transfected Flag-YAP-5SA with HA-YY1 by Co-IP. IP was performed using anti-YAP1 or anti-Flag antibodies and normal rabbit IgG antibodies. G IF staining was performed to determine the colocalization of YAP1 (red) and YY1 (green) in HCT116 and HT29 cells treated with CXCL12 (100 ng/ml) in the presence of AMD3100 (2 μM). DAPI was used for nuclear staining. Scale bars, 50 µm. (H) HCT116 cells were co-transfected with miR-124-3p or miR-188-5p promoter luciferase construct (miR-124 or miR-188 promoter) together with HA-YY1 plasmid. pGL3-basic plasmid was used as the vector control. The comparison of luciferase activities of promoter constructs normalized to Renilla activity was indicated. Bars are means ± SD; *P < 0.05, **P < 0.01 (n = 3)