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. 2022 Sep 15;13:5412. doi: 10.1038/s41467-022-33073-7

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© The Author(s) 2022

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. 6 — a The schematic of PGMLV-GC0643 construction. b Effect of GC0643 gene transfection on HGC27 cancer cells, as monitored by qRT-PCR of GC0643 mRNA expression fold change plot and western blot of anti-DDDDK-Tag. Data are presented as mean values ± SD from n = 3 biological replicates. Data were analyzed statistically by two-tailed Student’s t-test. ***P = 4.50E–05. c The mRNA expression of GC0643 was examined by RNAScope. The positive signals (red dots) were observed in normal mucosa, but reduced in cancer tissue (right) (scale bar represents 20 μm). Representative images from n = 3 biological replicates. d, e Cell growth activity (CCK8) and colony formation (soft agar assays) are presented. d Data are presented as mean values ± SD of 5 biological replicates, and were analyzed by two-tailed Student’s t-test with ^nsP = 0.547, **P = 0.002, ***P = 9.99E–07, ***P = 0.0008. e Data are presented as mean values ± SD of 3 biological replicates, and were analyzed by two-tailed Student’s t-test with **P = 0.0002. f The proliferating cell ratio examination by EdU (scale bar represents 20 μm). Representative images of EdU positive cells (red dots) from n = 5 biological replicates. g, h Enforced GC0643 induced cell apoptosis and resulted in G2/M arrest. Data are presented as mean values ± SD of 5 biological replicates. P values were derived from two-tailed Student’s t-test with *P = 0.01; ***P = 1.27E–05. i Cell migratory ability was suppressed by scratch wound healing assays. P values were derived from two-tailed Student’s t-test with *P = 0.02; **P = 0.08. Data are presented as mean values ± SD of 3 biological replicates. j The changed downstream pathways based on GC0643 overexpression. k The knockdown efficiency of shRNA for GC0643 overexpressed cancer cells. P values were derived from two-tailed Student’s t-test with ***P = 5.81E-06; ***P = 4.26E–06. Data are presented as mean values ± SD of 3 biological replicates. l, m Cell migration ability are suppressed, but reversed after knockdown GC0643 by shRNA. P values were derived from two-tailed Student’s t-test with ***P = 4.70E-05, ^nsP = 0.067, *P = 0.013. Data are presented as mean values ± SD of 4 biological replicates.