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. 2017 Mar 30;8(3):e2714. doi: 10.1038/cddis.2017.134

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Copyright © 2017 The Author(s)

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Foxi1 induces miR-491-5p promoter activity in gastric cancer cells. (a) The expression levels of Foxi1 mRNA in gastric cancer tissues were analyzed by qRT-PCR. (b) qRT-PCR analysis of Foxi1 expression in normal gastric mucosal and gastric cancer cells and normalized against U6 RNA. (c) Schematic diagram of the putative miR-491 promoter with one potential Foxi1 response element. (d) Luciferase activity of reporter constructs spanning the putative Foxi1-binding site or a negative control sequence. (e) ChIP assays were performed with control (rat IgG), anti-Foxi1 antibody to determine Foxi1 occupancy of miR-491 promoter. (f) qRT-PCR analysis was performed with primers spanning predicted Foxi1-binding site of miR-491 promoter. (g) The expression of miR-491-5p was assayed by qRT-PCR after transfection with Foxi1 vector or negative control in MKN45/SGC-7901 cells. (h) The expression of Wnt3a was determined by qRT-PCR after transfection with Foxi1 vector or negative control in gastric cancer cells. (i) MTT assay was performed to test the impact of gastric cancer cells treated with Foxi1 expression vector. (j) Colony formation was applied to measure the effect of gastric cancer cells treated with Foxi1 expression vector. (k) Apoptosis was conducted to investigate the impact of gastric cancer cells after transfection of Foxi1 expression vector. (l) Cell cycle distribution was analyzed by flow cytometry to determine the effect of gastric cancer cells treated with Foxi1 expression vector. (m) The expression of pro-caspase 3, active caspase 3, cleaved PARP, BCL-2, CDK6, and CCND1 was detected by western blot (*P<0.05, **P<0.01)