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. 2018 Aug 7;8:301. doi: 10.3389/fonc.2018.00301

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Copyright © 2018 Pelekanou, Notas, Athanasouli, Alexakis, Kiagiadaki, Peroulis, Kalyvianaki, Kampouri, Polioudaki, Theodoropoulos, Tsapis, Castanas and Kampa.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of APRIL and BAFF on mammosphere formation and breast cancer stem cell population and of testosterone on the effect of APRIL and BAFF on mammosphere formation (A). Percentage of primary mammospheres formed after treatment of breast cancer cells (T47D and MDA) with either BAFF or APRIL (100 ng/ml) for 9 days. Results of three independent experiments are expressed as a percentage of control (untreated) cells (mean ± SE, *denotes p < 0.05, at least). (B) Percentage of mammospheres (primary) formed after treatment of breast cancer cells (T47D and MDA) with either BAFF or APRIL (100ng/ml) in the presence or absence of testosterone-BSA, for 9 days. Results of three independent experiments are expressed as a percentage of control (untreated) cells performed (mean ± SE, *denotes p < 0.05). (C) Percentage of mammospheres formed after treatment of T47D cells with either BAFF or APRIL (100 ng/ml) for 9 days (primary mammospheres) compared to secondary mammospheres and a subsequent 7-day period. Results of three independent experiments are expressed as a percentage of control (untreated) cells. (mean ± SE, *p < 0.05) (D). Percentage of breast cancer stem cells (T47D and MDA) with or without treatment with BAFF or APRIL (100 ng/ml) for 4 days as estimated by assaying them with high green autofluorescence on a population of 20,000 cells, in a BL2-A/BL1-A dot plot. Three independent experiments were performed and the results of a representative one is presented. (E) Comparison of the percentages of breast cancer stem cells as estimated by autofluorescence on a population of 20,000 cells in a BL2-A/BL1-A dot plot and ALDEFLUOR kit with or without treatment with BAFF or APRIL (100 ng/ml) for 4 days. Three independent experiments were performed (mean ± SE, *denotes p < 0.05). (F) Comparison of the percentages of breast cancer stem cells (identified by autofluorescence on a population of 20,000 cells in a BL2-A/BL1-A dot plot) in T47D cells and mammospheres treated or not with BAFF or APRIL (100 ng/ml) for 4 and 12 days respectively. Three independent experiments were performed (mean ± SE, *denotes p < 0.05).