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. 2018 Oct 16;37:252. doi: 10.1186/s13046-018-0927-8

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© The Author(s). 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 1 — Expression of CSC markers and miRNA-302 family in LCSCs. qRT-PCR (a) and Western blot (b) to quantify CD133 and EpCAM level in adherent and tumor spheres of HepG2 and Huh7 cells after 3 days cultured in stem cell medium containing EGF and bFGF. GAPDH was used as loading control. c, MiRNAs microarray analysis to identify key miRNA molecules implicated in LCSCs maintenance. qRT-PCR to validate the miRNA-302a (d) and -302d (e) level in adherent and tumor spheres of HepG2 and Huh7 cells after 3 days cultured in stem cell medium containing EGF and bFGF. f, Western blot to quantify CD133 and EpCAM level in tumor spheres of HepG2 and Huh7 cells at Day 7 cultured in stem cell medium and differentiated liver cancer cells. qRT-PCR to quantify the miRNA-302a (g) and -302d (h) mRNA level in adherent and tumor spheres of HepG2 and Huh7 cells at Day 7 cultured in stem cell medium and differentiated liver cancer cells. Data shown are the means ± SD of three independent experiments. Statistical analyses were performed with one-way ANOVA (*P < 0.05, **P < 0.01, and ***P < 0.001 vs. normal)