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. 2018 Jan 24;7(1):7. doi: 10.1038/s41389-017-0022-6

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© The Author(s) 2018

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 Interaction between EBAG9 and TM9SF1 in 293T cells. Lysate of 293T cells transfected with Flag-EBAG9 and HA-TM9SF1 plasmids was immunoprecipitated with anti-EBAG9, anti-TM9SF1 or control IgG, then subjected to western blot analysis using EBAG9 (left panel) or TM9SF1 (right panel) antibody. Arrows show signals for Flag-EBAG9 and HA-TM9SF1. b Subcellular co-localizatiion of EBAG9 and TM9SF1. HeLa cells were transfected with GFP-TM9SF1 and DsRed-EBAG9, and subjected to fluorescent microscopic examination. Scale bars, 10 μm. c TM9SF1 silencing inhibits LNCaP cell migration. Cells transfected with siRNA targeting TM9SF1 (siTM9SF1 #1) or siControl were seeded on the upper chamber and migrated cells were stained after 48 h. Representative images are shown. Scale bar, 20 μm. Migrated cell numbers were counted in 5 microscopic fields at least. Data are shown as mean ± SD (n = 5). d TM9SF1 modulates EMT-related gene expression in LNCaP cells. qRT-PCR analyses for indicated genes were performed using RNAs prepared from LNCaP cells treated with siTM9SF1 #1 or siControl. e TM9SF1 contributes to cell migration interacting with EBAG9. LNCaP-EBAG9 #4 and LNCaP-Vector #5 cells were transfected with siTM9SF1 #1 or siControl, and cell migration was examined as described above. Data are shown as mean ± SD (n = 5). f TM9SF1 plays a role in EMT-related gene expression along with EBAG9. LNCaP-EBAG9 #4 and LNCaP-Vector #5 cells were transfected with siTM9SF1 #1 or siControl, and SNAI2 mRNA expression was quantified as described. Data are shown as mean ± SD (n = 3). *P < 0.05; **P < 0.01 (two-sided Student’s t-test)