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. 2022 Jan 5;11:708915. doi: 10.3389/fonc.2021.708915

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Copyright © 2022 Fertal, Zaidi, Stein, Stein and Heath

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 CXCR4 inhibition on cell proliferation. U937 cells are kept in co-culture with human mesenchymal stem cells (hMSCs) and the cytotoxic effect of a CXCR4 inhibitor, AMD3465 is examined. (A) The expression of CXCL12 and CXCR4 in hMSCs is shown, normalized to GAPDH. (B) AMD3465 produces a decrease in pERK/ERK in U937 cells kept in co-culture in regular media. (C) The impact of AMD3465 on pERK/ERK in U937 cells kept in co-culture in charcoal stripped, phenol-free media in the presence of CXCL12 is shown by western blot, with corresponding densitometry. (D) The cytotoxic effect of AMD3465 is examined, both with and without doxorubicin. (E) Representative FACS data is shown. Propidium iodide (PI) staining delineates live versus dead cells, and CD45 positivity separates the CD45+ leukemia cells from the CD45- hMSCs. (F) The anti-proliferative effect of CXCR4 inhibition is evaluated by BrdU assay, with and without treatment dose cytarabine. (G) The anti-proliferative effect of high dose and low dose CXCR4 inhibition is evaluated by BrdU assay, with and without subtoxic dose cytarabine.