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. 2019 Feb 11;38(23):4590–4604. doi: 10.1038/s41388-019-0743-4

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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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Fig. 3 — PKNOX2 suppressed GC cell proliferation via induction of apoptosis and cell cycle arrest. a PKNOX2 overexpression in AGS and MKN45 cells was confirmed by RT-PCR and Western blot. b Subcellular localization of PKNOX2 in AGS and MKN45 cells. c PKNOX2 overexpression suppressed cell proliferation (both P < 0.0001) and (d) colony formation(AGS, P < 0.05;MKN45,P < 0.001). e Knockdown of PKNOX2 in GES1 cells was determined by RT-PCR and Western blot. f PKNOX2 silencing promoted GES1 cell growth (P < 0.01)and colony formation (P < 0.05). g PKNOX2 induced apoptosis in AGS and MKN45 cells. Apoptosis was determined by flow cytometry after Annexin V-PI/7-AAD staining (left)(AGS, P = 0.0179;MKN45, P = 0.0002). Apoptosis markers activated by PKNOX2 were determined by Western blot (right). h PKNOX2 increased cell population in G₀/G₁ phase(P < 0.0001), together with decreased S phase population (left)(P < 0.0001). PKNOX2 inhibited the expression of cyclin D1 and CDK4; but induced p21 and p27 expression (right). (*P < 0.05; **P < 0.001; ***P < 0.0001)