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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. 1 — a Gene set enrichment analysis using the C6 canonical pathways Broad MsigDB database on gene expression data compared control and LCN2 knockdown MDA-MB-231 cell lines. b, c Calcein-AM analysis cellular iron level of MCF-7 cells transfected with indicated YAP mutant (b). Scale bar, 100 μm. Values were normalized to control group (c). (mean ± SD, n = 3, One-way ANOVA analysis). d Enhanced DAB iron staining (n = 69) and immunohistochemistry staining of YAP (n = 69) in breast cancer tissue arrays. Scale bar, 100 µm. e Correlations between the YAP and iron levels in human breast tumors were analyzed. two-sided chi-square test; R, correlation coefficient. f, g Schematic illustration of the analysis of lncRNA profiles stimulated with FAC (100 μM) or DFO (100 μM) from the human Lincode® siRNA library into MCF-7 cells that were engineered with a TEAD-driven luciferase reporter (f). The representative candidates were assayed by RT–qPCR (g) (mean ± SD, n = 3, One-way ANOVA analysis). h Calcein-AM analysis the cellular iron level of HEK-293T with candidate lncRNAs knockdown. Values were normalized to control group. (mean ± SD, n = 3, two-sided Student’s t test). (i) RT–qPCR detected the LncRIM expression in breast cancer tissues (n = 98) and paired adjacent tissues (n = 36). The horizontal black lines represent median values. (mean ± SD, two-sided Student’s t test). j Kaplan–Meier analysis of overall survival of breast cancer patients with low versus high expression of LncRIM (n = 98, Gehan–Breslow test). k, l Calcein-AM detected cellular iron level of control and LncRIM knockdown MCF-7 cells (k). Scale bar, 100 μm. Values were normalized to control group (l). (mean ± SD, n = 3, One-way ANOVA analysis). m Colony formation assay of control and LncRIM knockdown MCF-7 cells with or without 200 µM FAC stimulation (mean ± SD, n = 3, Two-way ANOVA analysis). n Colony formation assay of EV and LncRIM overexpressed MCF-7 cells treated with or without DFO (100 µM) (mean ± SD, n = 3, Two-way ANOVA analysis). o Graphical illustration of the main proteins involved in the cellular iron homeostasis. p RT-qPCR and Heatmap detected the expression of iron metabolism-related genes in control and LncRIM knockdown MCF-7 cells. (mean ± SD, n = 3, One-way ANOVA analysis). q RT–qPCR detection of LncRIM expression in cytoplasmic and nuclear fractions. (mean ± SD, n = 3).