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. 2023 Apr 20;14:2253. doi: 10.1038/s41467-023-37871-5

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© The Author(s) 2023

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. 3 — a, b The expression of DMT1 and TFR1, and the levels of p-LATS1 (S909, T1079), and p-YAP (S127) were detected in control, LncRIM knockdown or LncRIM overexpressed MCF-7 and MDA-MB-468 cells. c–e LncRIM promoted the expression of DMT1 and TFR1 in YAP-dependent manner. Immunoblot (c) and RT-qPCR were performed to detect the expression of DMT1 (d) and TFR1 (e) in control and YAP knockdown MCF-7 cells with or without overexpression of LncRIM. (mean ± SD, n = 3 biologically independent experiments, two-way ANOVA analysis). f The cellular iron level of control and YAP-silenced MCF-7 cells with or without LncRIM overexpression was measured by Calcein-AM assay. Values were normalized to the control group (mean ± SD, n = 3, Two-way ANOVA analysis). g, h The expression of DMT1 and TFR1 (g) as well as the cellular iron level (h) was detected by immunoblot and Calcein-AM assay in YAP-rescued MCF-7 cells. Values were normalized to those in the control group. (mean ± SD, n = 3, One-way ANOVA analysis). i MEME analysis of YAP/TEAD binding motif in the DMT1 promoter region by using GSE38369. j YAP/TEAD directly regulates the transcription of DMT1. Chromatin immunoprecipitation (ChIP) assay combined with RT-qPCR were performed by using the IgG and TEAD4 antibodies. (mean ± SD, n = 3, two-sided Student’s t test). k Luciferase reporter assay was performed with overexpression of YAP-5SA and DMT1-promoter or DMT1-promoter deleted mutants (CATTCT) in MCF-7 cells (mean ± SD, n = 3, Two-way ANOVA analysis). l, m Immunoblot (l) and Calcein-AM assays (m) were performed to assess the expression of DMT1, TFR1 and iron level in control or IRP2 knockdown MCF-7 cells with overexpression of YAP or LncRIM. (mean ± SD, n = 3, two-way ANOVA analysis).