The tco89-71 mutation partially restores phosphorylation of Ipl1 substrates. (A) Immunoblot analysis of WT (KT1113), glc7-127 (KT1969), glc7-127 ipl1-2 (KT1968), ipl1-2 (KT1829), tco89-71 ipl1-2 (KT3003), and tco89-71 (KT2959) extracts with phospho-specific antibody to histone H3 serine 10 and with antibody to total histone H3. (B) Dam1 phosphorylation on Ser257 of WT (KT1113), glc7-127 (KT1969), glc7-127 ipl1-2 (KT1968), ipl1-2 (KT1829), tco89-71 ipl1-2 (KT3003), and tco89-71 (KT2959) extracts was examined by immunoblotting with anti-S257 P and the overall level of Dam1 was examined using anti-Dam1. The ratio of the signal from the anti-S257 P antibody to the anti-Dam1 antibody is presented below each lane in A and B. (C) The fold increase in phosphorylation of H3 Ser10 and Dam1 Ser257 in extracts from ipl1-2 tco89-71 and ipl1-2 glc7-127 strains over extracts from ipl1-2 strains. The results represent the average of three separate experiments for histone H3 and four separate experiments for Dam1. Error bars, SD.