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. Author manuscript; available in PMC: 2010 May 1.
Published in final edited form as: Cell. 2009 May 1;137(3):459–471. doi: 10.1016/j.cell.2009.02.027

Figure 2. RAD6 Mediates Endogenous H2B Ubiquitylation and H3K4-H3K79 Methylation in Human Cells.

Figure 2

(A) Human RAD6 can fully complement the function of yeast Rad6 for H2B ubiquitylation and H3K4-H3K79 methylation. Yeast whole cell extracts from a wild-type strain (containing a chromosomal FLAG-H2B gene), its isogenic Δrad6 strain, and Δrad6 strains that harbor the indicated E2 expression plasmids (driven by the natural yeast Rad6 promoter) were subjected to immunoblotting with the indicated antibodies. Two different amounts of cell extracts were loaded. (B) Effects of siRNA-mediated knockdown of H2B ubiquitylation factors on histone modifications. 293T cells were treated with control, hBRE1A, hBRE1B, hRAD6A, hRAD6B and hUbcH6 siRNAs indicated. Total cell lysates were subjected to immunoblot analyses with indicated antibodies. A nonspecific band in the hUbcH6 immunoblot is indicated by an asterisk. (C) Total RNAs from siRNA-treated cells were subjected to RT-PCR analyses for mRNA levels of indicated genes.