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. 2020 Apr 20;11(4):248. doi: 10.1038/s41419-020-2471-7

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© The Author(s) 2020

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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Fig. 2 — The knockdown efficiency of siRNAs was determined by a RT-PCR and b western blotting. c MTT assays were performed to determine the proliferation of SMMC-7721 and BEL-7402 cells transfected with siRNAs. d The effects of silencing SPR on cell proliferation of SMMC-7721 and BEL-7402 cells were detected by colony formation assays. e RTCA assays were performed to evaluate the effect of SPR knockdown on HCC cell proliferation. f Cell apoptosis was measured by flow cytometry using annexin V/PI staining in cells after siSPR treatment. g Apoptosis in different groups was analyzed by PARP cleavage and Caspase 3 cleavage analysis using western blots. h Hoechst staining of the cellular nuclei was used to identify apoptosis in HCC cells. Scale bar: 100 μm. i The level of cytochrome C (Cyt C) in the cytoplasm was detected in SMMC-7721 and BEL-7402 cells transfected with siSPR. Data are represented as the mean ± SD of three independent experiments. The p-values < 0.05 were considered statistically significant for all tests.