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. 2018 Apr 12;13(4):e0194925. doi: 10.1371/journal.pone.0194925

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© 2018 Erdei et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 4 — Functional analysis of ABCG2 in EGFP-HUES9 (control) and GFP-ABCG2 expressing cells (A) Examination of the transport function of the GFP-ABCG2 protein variants in HUES9 cells by measuring DCV uptake. Cellular DCV fluorescence was measured in cells expressing the R482R (wild-type) and R482G (substrate mutant) variants of GFP-ABCG2. The purple histogram shows the signal of the DCV in the control cells and the green histogram shows the signal of DCV in the Ko143-treated cells. (B) Analysis of mitoxantrone cytotoxicity in EGFP-HUES9 (control) cells and in HUES9 cells expressing GFP-ABCG2 variants. Survival of parental and GFP-ABCG2 expressing HUES9 cells was measured in the presence of 5–200 nM Mitoxantrone (MX) by flow- cytometry. The ratio of the dead and living cells was calculated on the basis of propidium-iodide accumulation and was normalized to untreated cells. Values represent the means±S.D. of 3 independent experiments. Significant differences (Student’s t-test, P<0.01) in the survival of parental and ABCG2-variants expressing clones are indicated by asterisks. The exact mean±S.D. and p values are presented in S1 Table.