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. 2021 May 31;11:11321. doi: 10.1038/s41598-021-90887-z

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© The Author(s) 2021

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 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/.

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Expression and subcellular localization of maspin in non-tumorigenic and tumorigenic cell lines. (A) Relative mRNA expression level of maspin in MCF10A cells. mRNA expression was normalized to level of GAPDH. Data are shown as the mean ± SD (n = 3). (B) Protein expression level of maspin in MCF10A cells. GAPDH is shown for loading control. Whole western blots are presented in Supplementary Fig. S7. (C) Relative mRNA expression levels of maspin in breast cancer cell lines. mRNA expression was normalized to levels of GAPDH. Data are shown as the mean ± SD (n = 3). (D) Protein expression levels of maspin in three breast cancer cell lines. Expression level of GAPDH was used as a protein loading control. Whole western blots are presented in Supplementary Fig. S8. (E) Representative images of subcellular localization of maspin protein in MCF10A, MCF-7, and MDA-MB-231 cells. Expression of maspin protein was detected using anti-maspin antibody (clone G167-70). Nuclei was counterstained with DAPI. Scale bars represent 20 μm. Images were acquired using Olympus FLUOVIEW FV1000D confocal laser scanning microscope and Olympus FV10-ASW software (ver. 02.01). (F) Quantification of the subcellular localization of maspin. Bar graph shows the percentage of maspin distribution that is calculated by counting number of cells exhibiting pancellular, nuclear, or cytoplasmic distribution (n = 3). (G) Protein expression of maspin in the cytoplasmic (Cyt) and nuclear (Nuc) fractions. Expression levels of HSP90 and Lamin B1 was used as protein loading control in the cytoplasmic and nuclear fractions, respectively. Whole western blots are presented in Supplementary Fig. S9.