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. 2020 Jan 9;9:1431. doi: 10.3389/fonc.2019.01431

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Copyright © 2020 Cai, Bi, Bai, Zhang, Zhai, Xiao, Tang, Yang, Zhang, Li, Yang, Liu, Chen, Sun, Liu and Yang.

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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GA reduces stemness and induces differentiation of hepatic cancer cells. (A) Survival of HepG2 and PLC/PRF/5 cells incubated with the indicated amounts of GA for 48 h. (B) Cell proliferation was assessed through a colony formation assay. The number of colonies was compared (one-way ANOVA; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001). (C) Images of sphere formation assay of HepG2 and PLC/PRF/5. The number of spheres was compared (one-way ANOVA; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001). (D) Expression of representative CSC markers (SOX2 and OCT4) was analyzed by Western blot assay (one-way ANOVA; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001). (E) Images of HepG2 and PLC/PRF/5 cells were taken after incubation with different GA concentrations. (F) Expressions of representative differentiation markers (AFP and HEPPAR1) were analyzed by Western blot assay (one-way ANOVA; ^*P < 0.05, ^**P < 0.01). Scale bars in C and E are at 200 and 20 μm, respectively.