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. 2018 Jul 24;5:90. doi: 10.3389/fcvm.2018.00090

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Copyright © 2018 Puttini, Plaisance, Barile, Cervio, Milano, Marcato, Pedrazzini and Vassalli.

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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Cell viability, clonogenicity, cell growth, intracellular RA levels, effects of DEAB and all-trans RA. (A) Freshly isolated cells. Flow cytometry analysis of DAPI, Aldefluor (with and without DEAB), and Annexin V (right panel). Part of ALDH^lo cells are Annexin V-positive (K-+). (B) Freshly isolated ALDH^hi cells and unsorted cells. Clonogenic analysis (n = 3/group). (C) In vitro growth kinetics. Left panel: freshly isolated cells; right panel: outgrowth cells (n = 3/group in both panels). (D) Dose-dependent effects of DEAB on EdU incorporation and cell density. (E) RA concentrations in ALDH^hi and ALDH^lo cells measured by ELISA. (F) Dose-dependent effects of all-trans RA (ATRA) on cell density in the presence of DEAB (100 μM).