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. 2020 Oct 2;11(10):822. doi: 10.1038/s41419-020-03030-7

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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. 4 — a Western blotting showed the effects of AFP overexpression or knockdown on the expression of Fas/FasL/FADD. Left, HLE cells were stably transfected with AFP expression plasmids (AFP1# and AFP2#) or an empty vector (Vec). Right, HuH7 and HepG2 cells were transfected with a nontarget control (NC) or siRNAs against AFP (siAFP1# and siAFP2#). b Restoration of Fas expression counteracted AFP-enhanced cell growth. Fas was re-expressed in the HLE cells stably expressing AFP (AFP1# and AFP2#). Cell growth was determined by a clonogenicity assay. The expression levels of AFP and Fas/FADD were detected by Western blotting (left). ***P < 0.001. c The expression of AFP and Fas/FADD in AFP-positive wild-type (wt) and AFP-deficient (afp-/-) mouse liver tumors was determined. Left panel, representative bands of a Western blot are shown. Right panel, semiquantitative analysis of the Fas protein levels in AFP-positive wt (n = 20) and afp-/- (n = 17) mouse liver tumors is shown. Detailed Western blot data can be found in the Supplementary figures. ***P < 0.001 versus wt. d A Western blot shows the expression of AFP and Fas/FADD in human HCC specimens. e The protein expression of AFP and Fas in HCC patients was examined. A tissue microarray containing 95 primary human HCC specimens was used to evaluate the AFP and Fas protein levels by IHC, and representative images from three specimens are shown. Scale bar, 100 μm. f The correlation between the AFP and Fas proteins in HCC specimens was examined. Staining for AFP and Fas in the above tissue microarray was quantified, and the correlation between the AFP and Fas proteins was analyzed by the Pearson correlation test.