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. 2003 Dec 11;23(1):191–201. doi: 10.1038/sj.emboj.7600029

Figure 6.

Figure 6

Orc1p is released from pEPI-1 during the S phase in HeLa cells. A HeLa cell clone containing episomal pEPI-1 was investigated (see Figure 1B). (A) Left, FACS analysis showing the passage of cells through the S phase. Right, conventional PCR using DNA from crosslinked chromatin. PCR was performed with 5 ng nucleoprotein before immunoprecipitation (Input) or 1/100th of the recovered DNA after immunoprecipitation with Orc1p (α-Orc1), Orc2p (α-Orc2) and MCM3p (α-Mcm3) antibodies using primer pairs SV40 and pCMV with 20/23 cycles. (B–D) Quantitative real-time PCR were performed with six different primer pairs on DNA templates extracted from chromatin precipitated with antibodies against Orc1p, Orc2p or Mcm3p.