FIG. 2.

Chromatin association of Orp1 and Mcm proteins in synchronized cells. OGC9 (h⁻ cdc25-22 orp1::5xFLAG-orp1) cells were incubated at 36°C for 4 h to produce arrest at the G₂/M boundary and released at the permissive temperature (25°C). (A) Every 20 min after release, whole cell extract (WCE), the soluble fraction, and the insoluble chromatin fraction were analyzed by immunoblotting with anti-FLAG, anti-SpMcm6, or anti-SpMcm2 antibodies. The amounts of proteins were adjusted by immunoblotting with anti-tubulin antibody. The arrowheads indicate the positions of FLAG-Orp1p, SpMcm6p, and SpMcm2p. (B) The septation index is shown along with schematic illustrations of corresponding phases of the cell cycle.