FIG. 4.

Spt6-dependent H3K36me3, but not H3K36me2, when Set2 protein is restored. (A) Spt6 influences the levels of Set2 protein. Western blots of total protein extracts from the strains indicated incubated with the antibodies shown. (B) Western blots of total protein extracts from strains expressing SET2 from the GAL1 promoter integrated at the SET2 locus in the FY2181 background (WT or spt6-1004) cultured in raffinose (lane 2; YPR off) or galactose (lane 3; YPG on) incubated with the indicated antibodies. Lane 1 contains extract from the WT parental background in which SET2 is expressed from the endogenous SET2 promoter. (C) Chromatin immunoprecipitation to detect Set2 at three positions on STE11, the coding region of MET16 (CR), and telomere 01L in the strains indicated, cultured at 30°C in raffinose (YPR off) or galactose (YPG on). Data are expressed as a percentage of the input signal. Note that the profile for Set2 at STE11 detected using the polyclonal antibodies is different from that with the FLAG epitope (see Fig. 1D). These differences are reproducible and have been observed by others using different epitope tags on Set2 (7).