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. 2014 Apr 29;42(11):6901–6920. doi: 10.1093/nar/gku312

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — ASH2L is required for efficient gene expression and H3K4 trimethylation. (A) HEK293T cells were transiently transfected with siRNA oligo pools targeting ASH2L or MYC mRNA or a control oligo pool (Ctrl). Expression of ASH2L, MYC, CCND2, ODC and NCL mRNA was analyzed by quantitative RT-PCR and normalized to GUS. Error bars represent s.d. (n = 4). (B) Cell lysates of HEK293T cells from an experiment shown in panel A were used for western blot analyses with the indicated antibodies and actin as loading control. (C)–(H) HEK293T cells were transiently transfected with siRNA pools targeting ASH2L mRNA. Subsequent ChIP experiments were performed with antibodies against ASH2L (C), H3K4me3 (D), RbBP5 (E), WDR5 (F), MYC (G) and H3ac (H). DNA fragments were analyzed as described in the figures before. Error bars represent s.d. of PCR triplicates.