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. 2024 Dec 28;14:31210. doi: 10.1038/s41598-024-82514-4

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 3 — LF induces cell apoptosis through the activation of p38 MAPK and JNK signaling. (A) Western blot images showing the entry of LF and the effects of LF treatment for 48 h on p38 MAPK, JNK, and ERK phosphorylation. The relative phosphorylated profiles shown below were determined by normalizing the phosphorylated protein intensity to the unphosphorylated protein intensity and comparing them with the DPBS-treated HepG2 lysates. ND means that the band intensity was not detectable. (B) Western blot images showing the time-course of p38 MAPK, JNK, and ERK phosphorylation after 5 mg/ml LF treatment. (C) Representative cell images at 24 and 48 h after treatment with LF (5 mg/ml) and inhibitors against p38 MAPK (SB203580) and JNK (SP600125). Scale bars, 100 μm. (D) Cell viability after treatment with LF and inhibitors (*, vs. no LF; #, vs. LF/no inhibitors). (E) Representative apoptosis analysis after treatment with LF and inhibitors. (F) The proportions of apoptotic and necrotic HepG2 cells after treatments with LF and inhibitors (*, vs. LF/no inhibitors). (G) Western blot images showing the changes in p38 MAPK and JNK phosphorylation and the release of cytochrome C and cleaved caspase 3 after treatment with LF and inhibitors. The relative protein levels were determined after β-actin normalization. Statistical significance: * (p < 0.05), ** (p < 0.01), and *** (p < 0.001); ^# (p < 0.05), ^## (p < 0.01), and ^### (p < 0.001).