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. 2004 May 1;18(9):981–991. doi: 10.1101/gad.1202304

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Figure 9. — The Orc6 RXL motif helps protect cells from re-replicating. (A) ORC6-ps,rxl MCM7-NLS GAL-CDC6Δ2-48-HA cells can be induced to re-replicate. ORC6 wild-type and mutant strains containing MCM7-NLS and GAL-CDC6Δ2-48-HA were arrested with 15 μg/mL nocodazole in YEP/2% raffinose, and then Cdc6Δ2-48-HA was induced with 2% galactose. (B) ORC2-ps exacerbates the re-replication phenotype (methods same as in A). (C) Pre-RCs form on the chromatin in the ORC6-ps,rxl MCM7-NLS GAL-CDC6Δ2-48-HA re-replicating cells. ORC6-ps,rxl MCM7-NLS cells with or without GAL-CDC6Δ2-48-HA were arrested as above, and then the chromatin pellet was isolated and the DNAse I- and salt-sensitive proteins were extracted and visualized by Western blotting.