Figure 5.

Kdm4d associates with replication origins depending on components of pre-replicative complex. (A) Kdm4d and Mcm6 bound replication origins at G1 phase. HeLa cells were synchronized by mimosine (G1) or nocodazole (G2/M). Chromatin immunoprecipitation (ChIP) and quantitative polymerase chain reaction (qPCR) analysis using antibodies against Kdm4d or Mcm6 was performed. ChIP DNA was analyzed by real-time PCR using PCR primers amplifying indicated origins and flanking regions. The ChIP enrichments from two independent experiments were represented by mean ± SD. (B) Depletion of Orc2 or Mcm3 abolished replication origin binding of Kdm4d. ChIP-qPCR analysis using antibodies against Kdm4d or Mcm6 was performed on Orc2- or Mcm3-depleted HeLa cells. ChIP DNA was analyzed by real-time PCR using PCR primers amplifying indicated origins and flanking regions. The ChIP enrichments from two independent experiments were represented by mean ± SD.