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. 2012 Dec 21;7(3):452–463. doi: 10.1016/j.molonc.2012.11.004

Figure 3.

Figure 3

Identification of the actors of the epigenetic regulation of the NY‐ESO1 gene. (A) Impact of siRNA targeting Dnmt1, Dnmt3b, HDAC1 and G9a on the expression level of NY‐ESO1mRNA. None siRNA treatment induced the NY‐ESO1mRNA expression (all p > 0.05, t test). Mesenchymal stem cell (MSC) was used as positive control i.e. cells expressing NY‐ESO1mRNA. siRNA‐A is an aspecific siRNA used in control according to the manufacturer's instruction. (B–C–D). Sequential chromatin immunoprecipitation (reChIP) were performed by using indicated antibodies. GFP antibody was used as negative control for non‐specific ChIP. Only the relative enrichment obtained with the antibody directed against HDAC1 and Egr1 is significantly superior to the enrichment obtained with the GFP antibody (p < 0.05). (E–F–G). Immunoprecipitation (IP) monitoring the proteins interacting and co‐recruited with Dnmt1, Egr1 and HDAC1 respectively. NE: nuclear extract (positive controls for western blot analysis), GFP antibody was used to perform non‐specific IP (negative controls). Only the relative enrichment obtained with the antibody directed against Dnmt3b, mSin3A, NCOR1 and Egr1 is significantly superior to the enrichment obtained with the GFP antibody (p < 0.05, t test).