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Overproduction of Mcb1 causes dissociation of chromatin-bound MCM proteins. nmt1-cdc18 mcm2V5 mcm4GFP (FY4958), nmt1-mcb1HA mcm2V5 mcm4GFP (FY4961), and wild-type (FY11) overnight cultures were grown in low thiamine, then inoculated into −thiamine (−T) or +thiamine (+T) medium, grown at 32 °C for 14 h, and harvested for an in situ chromatin binding assay. Mcb1HA and Mcm2V5 localization was detected in untreated cells and Triton-treated cells with specific antibodies. Scale bar, 10 μm.
