Fig. 7

Both HMO1 and HMO1 deleted for its C-terminal tail compete with H1 for binding to chromatin. a Quantification by qRT-PCR of ChIP using antibody to H1 with DDY3, hmo1∆, and hmo1-AB strains, monitoring binding at the MAT locus. b qRT-PCR analysis of ChIP using antibody to H1, monitoring binding at 18S rDNA and at KRE5. Data were normalized to corresponding input control at each time point. Three independent experiments were performed. Error bars represent standard deviation. Asterisks represent statistical significance from DDY3 based on Student’s t test (P < 0.05). c Western blot using antibody to H1 showing equal protein level of histone H1 after transforming plasmid expressing human H1 under control of a strong, constitutive promoter in DDY3 (DDY3 H1), hmo1∆ (hmo1∆ H1), and hmo1-AB strain (hmo1-AB H1). Non-transformed cells DDY3, hmo1∆, and hmo1-AB were used as negative control. GAPDH expression levels were assessed in all samples as internal loading control, and the blots are representative of four independent experiments. GAPDH migrates with a Mw ~36 kDa, while H1 migrates with a Mw ~30 kDa (slower than its calculated Mw ~22 kDa). d Densitometric analysis of three separate blots from three independent experiments shown in (c). Relative H1 level = H1/GAPDH. Error bars represent standard deviation