Cells bearing a cds1+-HA allele were treated with 8 mM HU for ~3 h for activation of Cds1 kinase activity and subsequently harvested by centrifugation. The cell pellet was resuspended in lysis buffer (pH 7.6), phosphorylation inhibitor cocktail ,0.1% NP-40, and 1× proteinase inhibitor cocktail and subjected to a bead ruptor (bead beater). 

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