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. 2016 Jul 25;36(7):942–955. doi: 10.1038/onc.2016.260

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Copyright © 2017 The Author(s)

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 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-nc-sa/4.0/

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Identification of the Snail/HOTAIR/EZH2 complex. (a) RIP assays with rabbit polyclonal anti-Snail, anti-EZH2 or preimmune IgG on 24 h TGFβ-treated (TGFβ) or untreated (NT) murine hepatocyte cell extracts. RNA levels in immunoprecipitates were determined by qRT–PCR. HOTAIR lncRNA and, as controls, ribosomal L34 RNA, GAPDH pre-mRNA and SRA lncRNA were reported as percentage with respect to 1/10th of the input sample (TGFβ treated in the hotair panel or NT in the other panels) (% Input). Data are means ±s.e.m. of three independent experiments. (b) Co-immunoprecipitation of Snail and EZH2. Immunoprecipitations with rabbit polyclonal anti-Snail, anti-EZH2, preimmune IgG or no antibody (NoAb) were performed on protein extracts from murine hepatocyte cells treated with TGFβ for 24 h and silenced for HOTAIR (siHotair) or GFP (siCtr) as control. Immunoblots were performed using anti-Snail and anti-EZH2 antibodies. (c) RIP assays with rabbit polyclonal anti-Snail, anti-EZH2 or preimmune IgG on TGFβ-treated (TGFβ) or untreated (NT) human hepatoma cell extracts. RNA levels in immunoprecipitates were determined by RT–qPCR. HOTAIR lncRNA and, as controls, ribosomal L34 RNA, GAPDH pre-mRNA and SRA lncRNA were reported as percentage with respect to 1/10th of the input sample (TGFβ treated in the hotair panel or NT in the other panels) (% Input). Data are means ± s.e.m. of three independent experiments. (d) Co-immunoprecipitation of Snail and EZH2. Immunoprecipitations with rabbit polyclonal anti-Snail, anti-EZH2 or preimmune IgG were performed on protein extracts from human hepatoma cells treated with TGFβ for 24 h and silenced for HOTAIR (siHotair) or GFP (siCtr) as control. Immunoblots were performed using anti-Snail and anti-EZH2 antibodies.