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. 2021 Mar 5;40(14):2509–2523. doi: 10.1038/s41388-021-01711-x

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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 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 The Claudin low (CL)-TNBC subgroup (72 cases) showed an average age at diagnosis of 61.9 years, while the overall survival average was 126.5 months. The median months' survival calculated for the whole follow-up was 27.96 months. In total, 12/72 cases (17%) exhibited AURKA alterations (mRNA up-regulation and/or copy number variations), while none of 72 patients harbored deletions or down-regulations of AURKA. Survival analysis showed that aberrant AURKA expression was significantly associated with reduced patient overall survival (p-value = 0.00584). b Immunoblot assay showing expression of total and phosphorylated AURKA in BT-549 cells before and after treatment with TGF-β1 (10 ng/ml) for 48 h. c Densitometry analysis showing the fold change of total and phosphorylated AURKA protein levels in BT-549 TNBC cells normalized to α-Tubulin. Experiments were performed in triplicate. d BT-549 cells were treated with TGF-β1 (10 ng/ml) for 48 h. After 48 h incubation, ALDH1 activity was detected with Aldefluor kit and measured by FACS analysis on 10,000 events. ALDH1 inhibitor DEAB was used as control. Graph showing the average of ALDH1^High cells from three independent experiments (±s.d.). e Immunoblot assay showing expression of total AURKA in BT-549 cells infected with scrambled and AURKA lenti-shRNAs for 48 h. Densitometry analysis showing the fold change of total AURKA protein levels in BT-549 TNBC cells normalized to α-Tubulin. f BT-549 cells were treated with 10 ng/ml TGF-β1 and lenti-shRNAs. After 48 h incubation, ALDH1 activity was detected with Aldefluor kit and measured by FACS analysis on 10,000 events. ALDH1 inhibitor DEAB was used as the control for each sample. Graph showing the average of ALDH1^High cells from three independent experiments (±s.d.). g BT-549 cells were treated with 10 ng/ml TGF-β1, scrambled lenti-shRNAs, and lenti-shRNAs targeting SMAD3. After 48 h incubation, ALDH1 activity was detected with Aldefluor kit and measured by FACS analysis on 10,000 events. ALDH1 inhibitor DEAB was used as the control for each sample. Graph showing the average of ALDH1^High cells from three independent experiments (±s.d.).