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. 2019 Nov 14;9:16802. doi: 10.1038/s41598-019-53191-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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TIPRL is an essential element for liver cancer cell survival and stemness. Huh7 (a,f,g,j,k) and SK-Hep-1 (b,h,i) cells were transfected with siCont/siTIPRL. After 72 hours, MTT analyses were performed to determine both the proliferation index (a,b left) and survival ratio (a,b right) of the cells. (c–e) Spearman correlation was used to determine correlations between levels of TIPRL, LC3 and CD133 in HCC tissues. Each dot represents a single sample. (f–i) 72 hours after siTIPRL transfection, quantitative RT-PCR analyses were performed to determine the mRNA levels of the indicated genes using primers (Supplementary Table 3). (j) Immunocytochemistry was performed using an anti-ALDH antibody to determine the level of ALDH activity in siCont/siTIPRL-transfected Huh7 cells. For nucleus staining, DAPI was used, and scar bar, 50 μm. (k) Quantification of ALDH activity observed in (j). All experiments were independently repeated four times. Statistical differences (^*P < 0.05; **P < 0.01; ***P < 0.0001) were determined by 2way ANOVA (a,b), paired t-test (f–i), and an unpaired t-test (k).