Jhd2 and Gis1 demethylate H3K4 and H3K36, respectively, within the rDNA regions in vivo. (A) The levels of histone H3K4 methylation were analyzed using ChIP for the indicated strains. Antibodies against H3K4me3 or H3 were used. (B) Quantitation of the ChIP results in (A). The results for methyl-H3 were normalized to the total H3 signal and presented as fold enrichment relative to WT. Error bars indicate the SD from three PCRs performed using two independent chromatin preparations. (C) The levels of H3K4 and H3K36 methylation were analyzed by ChIP as shown in Figure 2B. Chromatin fractions were obtained from WT (BY4741) strains containing pRS325-GALpro, pRS325-GALpro-Jhd2-HA or pRS325-GALpro-Gis1-HA. Cells were grown on SC medium containing 2% galactose, followed by immunoprecipitation with the indicated antibodies. (D) Quantitation of the ChIP results shown in (C). The results were normalized to an internal control as shown in Figure 2C and to the total H3 signal. Error bars indicate the SD from three PCRs performed using two independent chromatin preparations, and asterisks indicate statistically significant differences compared with WT (*, P <0.05; **, P <0.01). IP, immunoprecipitation; WT, wild type.