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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 by targeting JNK1. (A) JNK1 expression in HepG2 and PLC/PRF/5 cells after being transfected with siNC and siJNK1 plasmids. Colony formation assay (B) and sphere formation assay (C) in HepG2 and PLC/PRF/5 cells transfected with siNC and siJNK1, followed by treatment with DMSO or GA (2 mM) for 48 h. The numbers of colonies and spheres were compared (one-way ANOVA; ^***P < 0.001). (D). Expressions of representative CSC markers (SOX2 and OCT4) were analyzed by Western blot assay (one-way ANOVA; ^***P < 0.01). (E) Images of HepG2 and PLC/PRF/5 cells transfected with siNC and siJNK1 followed by incubation with DMSO or GA (2 mM) for 48 h separately. (F) Representative markers of differentiation (AFP and HEPPAR1) were analyzed by Western blot assay (one-way ANOVA; ^*P < 0.05, ^***P < 0.001). Scale bars in (C,E) are at 200 and 20 μm, respectively.