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. 2020 Jun 18;10(6):925. doi: 10.3390/biom10060925

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Caudatin reduces YAP nuclear localization and regulates breast CSC formation. (A) The expression level of YAP in the total protein of MDA-MB-231-derived mammospheres was measured after treatment of the mammospheres with caudatin for 48 h using western blot analysis. (B) The transcriptional expression of the YAP gene was detected in mammospheres by real-time RT-qPCR using specific primers. β-actin was used as an internal control. (C) Immunofluorescence (IF) analysis of YAP (red) expression and localization in MDA-MB-231 cells under caudatin treatment was performed. (D) The expression level of YAP in the cytosolic and nuclear protein fractions of MDA-MB-231-derived mammospheres was measured after treatment of the mammospheres with caudatin using western blot analysis. (E) The expression levels of GR and YAP in MDA-MB-231 cells were measured after treatment with a siRNA specific for GR. (F) The effect of knocking down YAP expression using YAP-specific siRNA on mammosphere formation was evaluated. (G) The effect of verteporfin, a suppressor of the YAP/TEAD complex, on mammosphere formation was evaluated. (H–J) After treatment with caudatin, siRNA targeting GR or verteporfin, the transcriptional expression of the CYR61 and CTGF genes was detected by real-time RT-qPCR using specific primers. β-actin was used as an internal control. The data are presented as the mean ± SD of three independent experiments. ** p < 0.01 and *** p < 0.001 versus the DMSO-treated control group.

Caudatin reduces YAP nuclear localization and regulates breast CSC formation. (A) The expression level of YAP in the total protein of MDA-MB-231-derived mammospheres was measured after treatment of the mammospheres with caudatin for 48 h using western blot analysis. (B) The transcriptional expression of the YAP gene was detected in mammospheres by real-time RT-qPCR using specific primers. β-actin was used as an internal control. (C) Immunofluorescence (IF) analysis of YAP (red) expression and localization in MDA-MB-231 cells under caudatin treatment was performed. (D) The expression level of YAP in the cytosolic and nuclear protein fractions of MDA-MB-231-derived mammospheres was measured after treatment of the mammospheres with caudatin using western blot analysis. (E) The expression levels of GR and YAP in MDA-MB-231 cells were measured after treatment with a siRNA specific for GR. (F) The effect of knocking down YAP expression using YAP-specific siRNA on mammosphere formation was evaluated. (G) The effect of verteporfin, a suppressor of the YAP/TEAD complex, on mammosphere formation was evaluated. (H–J) After treatment with caudatin, siRNA targeting GR or verteporfin, the transcriptional expression of the CYR61 and CTGF genes was detected by real-time RT-qPCR using specific primers. β-actin was used as an internal control. The data are presented as the mean ± SD of three independent experiments. ** p < 0.01 and *** p < 0.001 versus the DMSO-treated control group.