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. 2010 Nov 5;9(21):4351–4363. doi: 10.4161/cc.9.21.13596

Figure 1.

Figure 1

Figure 1

HDAC11 binds origins in S phase, inhibits Cdt1-induced re-replication and suppresses MCM loading. (A) Synchronized HaCaT cells (verified by BrdU incorporation, top) were subjected to anti-HDAC11 ChIP and qPCR analysis at the indicated times for interactions to origin and non-origin sequences. (B and C) Cdt1 alone or Cdt1 plus HDAC11 was expressed in HeLa cells for 48 h with adenoviruses and subjected to flow cytometric analysis. Relative amounts of each virus and the percentage of cells with >4 N DNA are indicated. (D) Mcm2 chromatin binding was assessed by IB in HeLa cells infected with HDAC11 or control (GFP) viruses for 24 h. (E) Synchronized CHO cells were separated into soluble and chromatin fractions at the indicated times and subjected to IB. BrdU verified synchrony (data not shown). (F) Graphical representation of IB results from (E). (G) Indicated proteins (top) were expressed in 293T cells and subjected to IP and IB analysis indicated on right. IP and IB analyses were performed with anti-tag antibodies. (H) Flag-HDAC11, HA-Geminin and Myc-Cdt1 were co-expressed in 293T cells followed by anti-Flag purification. Eluates were separated on a size-exclusion column and analyzed by IB. The data in (G and H) are representative of three independent experiments with similar results.