Figure 4.

Gwl promotes checkpoint recovery. (A) As in Figure 3A, biotinylated dA-dT oligos bound to M-280 streptavidin beads were added to extracts for 30 min to activate the DNA damage checkpoint and then removed with a magnet to allow recovery. As indicated, these extracts had also been either mock-treated with Protein G Dynabeads or depleted of Gwl with anti-Gwl antibody bound to Protein G Dynabeads as described in Materials and Methods, or Gwl-depleted and then reconstituted with purified Gwl. Recovery in the extracts was then analyzed at 90 min by western blotting with the indicated antibodies. (B) As in (A) extracts with or without depletion of Gwl were monitored for Cdk activation by western blotting with the indicated antibodies at various time points during recovery.