Figure 4.

Plx1 is not required for Greatwall-induced mitotic entry. Cycling extracts were subjected to mock depletion or Plx1 depletion, and then incubated starting at t = 0 for 40 min at 23°C. The extract was then supplemented with buffer (PBS) or with WT or KD Greatwall purified from OA-treated Sf9 cells. One-microliter aliquots of extracts were subsequently collected every 10 min for immunoblot analysis. As expected, Plx1 depletion causes a defect in mitotic entry. However, Plx1 depletion does not interfere with Greatwall-induced mitotic entry even though Cdc25C does not obtain its full complement of M phase phosphorylations.