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. 2021 May 28;12(6):552. doi: 10.1038/s41419-021-03837-y

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© The Author(s) 2021

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 — A, B The construction of Hep3B PNO1- downregulation (sh-PNO1) and HLE PNO1-upregulation (PNO1) cell. C GSEA validating the correlation between PNO1 expression and the cell proliferation genes. D, E CCK-8 assay analysis of the impact of PNO1 knockdown or overexpression on Hep3B and HLE cell growth. F, G Colony formation assay showing the effects of PNO1 knockdown (F) or overexpression (G) on Hep3B and HLE cell growth. H, I Apoptotic rate was measured using annexin V/PI double staining in PNO1 knockdown (H) or overexpression (I) cells. J TEM analysis of cell ultrastructural characteristics in Hep3B sh-PNO1 and sh-Ctrl cells. Chromatin condensation and nuclear fragmentation are indicated by arrows. Scale bar = 20 μm. K Western blotting analysis of apoptosis-related protein levels in cells as in (A) and (B). L, M Quantification of relative protein expression. N, O Apoptosis was also evaluated by TUNEL staining (N) and quantification of TUNEL positive cells (O). Scale bar: 50 μm. Data were presented as mean ± SEM. n = 3–4. *p < 0.05, **p < 0.01, ***p < 0.001.