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. 2020 Sep 10;35(1):1697–1711. doi: 10.1080/14756366.2020.1818738

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© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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graphic file with name IENZ_A_1818738_F0003a_B.jpg — Representative flow cytometry dot-plots for Annexin V‐FITC assay in MCF-7 (A) and MDA-MB-231 cells (C) incubated with Epo (100 IU/ml) and LFM-A13 (100 μM) for 48 h (mean ± SD; n = 3). The live cells appear at the lower left corner in the plots; the early apoptotic cells appear at the lower right corner; the necrotic cells appear at the upper left corner; and dead cells appear at the upper right corner. The percentage of apoptotic MCF-7 cells (B) incubated with Epo and LFM-A13 is shown in the bar diagram as mean ± SD (n = 3). The percentage of apoptotic MDA-MB-231 cells (D) incubated with Epo and LFM-A13 is shown in the bar diagram as mean ± SD (n = 3). Immunoblotting analysis for Active caspase 6 and pro-caspase 6 in MCF-7 and MDA-MB-231 cells treated with erythropoietin beta (Epo 100 IU/ml), LFM-A13 (LFM 100 µM), and both for 48 h. Samples used for electrophoresis consisted of 20 µg of protein from 6 pooled cell extracts (n = 6). The band staining was quantified by densitometry. Bands of phospho-proteins were normalised to respective total proteins (E).

graphic file with name IENZ_A_1818738_F0003b_B.jpg — Representative flow cytometry dot-plots for Annexin V‐FITC assay in MCF-7 (A) and MDA-MB-231 cells (C) incubated with Epo (100 IU/ml) and LFM-A13 (100 μM) for 48 h (mean ± SD; n = 3). The live cells appear at the lower left corner in the plots; the early apoptotic cells appear at the lower right corner; the necrotic cells appear at the upper left corner; and dead cells appear at the upper right corner. The percentage of apoptotic MCF-7 cells (B) incubated with Epo and LFM-A13 is shown in the bar diagram as mean ± SD (n = 3). The percentage of apoptotic MDA-MB-231 cells (D) incubated with Epo and LFM-A13 is shown in the bar diagram as mean ± SD (n = 3). Immunoblotting analysis for Active caspase 6 and pro-caspase 6 in MCF-7 and MDA-MB-231 cells treated with erythropoietin beta (Epo 100 IU/ml), LFM-A13 (LFM 100 µM), and both for 48 h. Samples used for electrophoresis consisted of 20 µg of protein from 6 pooled cell extracts (n = 6). The band staining was quantified by densitometry. Bands of phospho-proteins were normalised to respective total proteins (E).