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. 2007 Dec 20;27(2):406–420. doi: 10.1038/sj.emboj.7601967

Figure 4.

Figure 4

Setd2 is a non-redundant H3K36-specific trimethyltransferase. (A) Cells were untransfected (−), mock transfected (no siRNA, mock) or transfected with Setd2 (0 and 3, two different siRNAs), NSD1 or non-targeting (non-t) siRNAs. Total mRNA was isolated 24 h later and relative levels of Setd2 and NSD1 mRNA were quantified by qRT–PCR and normalised to gapdh mRNA. A representative experiment is shown. Error bars represent the s.d. from triplicate PCRs. (B) Cells were transfected as in (A) and crude nuclear lysates were prepared 48 h later. Lysates were separated by 8% SDS–PAGE and transferred to PVDF membrane for immunoblotting with two different Setd2 antibodies (N-Setd2 (i) and C-Setd2 (ii)) and an MLL1 antibody (iii) as a control. (C) Crude nuclear lysates were prepared as in (B), separated by 15% SDS–PAGE and either Coomassie-stained (viii) or transferred to PVDF membrane for immunoblotting with various methyl (i–vi) or acetyl (ix–xv) modification-specific histone antibodies, as indicated in the figure. H4ac is an anti-histone H4 pan-acetyl antibody. Unmodified H3 (H3, vii) was used as a loading control.