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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. 4 — a Immunoblot was performed to examine the level of p-YAP (S127) and p-LATS1(S909, T1079). Serum-starved MCF-7 cells were treated with FAC (200 µM) or DFO (100 µM) for 24 h. b Serum-starved MFC7 cells were stimulated with different concentrations of FAC for 24 h. Immunoblot was performed to detect the level of p-YAP (S127) and p-LATS1(S909, T1079). c RT–qPCR detection of the LncRIM expression of MCF-7 cells stimulated with different concentration of FAC for 24 h. (mean ± SD, n = 3, one-way ANOVA analysis). d The expression of LncRIM in MCF-7 cells stimulated with different concentration of DFO for 24 h was analyzed with RT-qPCR. (mean ± SD, n = 3, one-way ANOVA analysis). e Luciferase reporter assay was performed of HEK-293T cells with overexpression of YAP-5SA and LncRIM promoter. (mean ± SD, n = 3, one-way ANOVA analysis). f YAP/TEAD4 directly regulates the transcription of LncRIM. CHIP-qPCR assay was performed by using IgG and TEAD4 antibodies. (mean ± SD, n = 3, two-sided Student’s t test). g RIP and RT–qPCR assays were performed to assess the interaction between LncRIM and NF2 of MCF-7 cells with or without FAC (200 µM) stimulation for 24 h. NF2 was immunoprecipitated by NF2 antibody and Protein A/G beads. IgG was used as the negative control. (mean ± SD, n = 3, One-way ANOVA analysis).(h) SFB-LATS1, HA-NF2 and LncRIM were co-transfected into MCF-7 cells with or without FAC (200 µM) stimulation for 24 h. SFB-LATS1 was immunoprecipitated by Flag beads. i, j Serum-starved control, LncRIM knockdown (i) or LncRIM overexpressed (j) MCF-7 and MDA-MB-468 cells were treated with FAC (200 µM) for 24 h, and the levels of p-LATS1(S909, T1079), p-YAP (S127) and FTH1 were detected by immunoblot. k, l Immunofluorescence staining of YAP in control and LncRIM knocked down serum-starved MCF-7 cells with or without FAC (200 μM) stimulation for 24 h (k). Scale bar, 10 µm. YAP localization in cells from three randomly selected fields of view was quantified (l). (mean ± SD, n = 3).