Figure 4.

Immunoprecipitation complexes formed after the simultaneous in vitro transcription/translation of the six spORC subunits. In vitro transcription/translation products derived from plasmids encoding all six spORC subunits (lane 1) were incubated on ice either without (lanes 2–5) or with DNase I (1,000 units/ml) (lanes 6–9). Complex formation was checked by immunoprecipitation (indicated as IP) with antibodies against Orc2p (lanes 2 and 6), Orc4p (anti-T7, lanes 3 and 7), Orc5p (HA, lanes 4 and 8), and Orc6p (lanes 5 and 9). After washing, the immunoprecipitated materials were subjected to SDS/PAGE and then autoradiographed.