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. 2021 Oct 9;12(10):924. doi: 10.1038/s41419-021-04234-1

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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. 4 — a The expression level of NME2 protein in NME2-silenced or -rescued in gastric cancer stem-like cells sorted from solid tumors of two gastric cancer patients. The protein was examined by western blot. β-Tubulin was used as a control. b Impact of NME2 knockdown and rescue on the cell number of gastric cancer stem-like cells. The gastric cancer stem-like cells sorted from solid tumors were transfected with NME2-shRNA to silence the NME2 expression. To rescue the NME2 expression, the NME2-silenced cells were transfected with the plasmid expressing NME2. At different days after transfection, the cell number was counted (^**p < 0.01). c Influence of NME2 knockdown and rescue on the cell viability of gastric cancer stem-like cells. The NME2-silenced cells were transfected with the plasmid expressing NME2, and the cell viability was examined at different times after transfection (^**p < 0.01). d Role of NME2 in cell cycle of gastric cancer stem-like cells sorted from solid tumors of two gastric cancer patients. The NME2-knock-down cells were transfected with the plasmid expressing NME2. At 48 h after transfection, the cells were subjected to the examination of cell cycle (^**p < 0.01). e Impact of NME2 knockdown and rescue on the caspase 3/7 activity of gastric cancer stem-like cells. The NME2-knocked-down cells were transfected with the plasmid expressing NME2. At 48 h after transfection, the caspase 3/7 activity of cells was examined (^**p < 0.01). f Flow cytometry detection of the impact of NME2 knockdown and rescue on apoptosis of gastric cancer stem-like cells sorted from solid tumors. The cells transfected with the plasmid expressing NME2 were cultured for 48 h, followed by the examination of apoptosis using Annexin V (^**p < 0.01). g Tumorsphere formation assay of the NME2-silenced or -rescued gastric cancer stem-like cells sorted from solid tumors. At day 8 after the inoculation of a single cell, the percentage of tumorsphere formation was calculated (^**p < 0.01). h Influence of NME2 knockdown and rescue on the expressions of stemness genes in gastric cancer stem-like cells from solid tumors. At 48 h after NME2 rescue, the expressions of stemness genes were examined with quantitative real-time PCR or western blot (^**p < 0.01).