Fig. 7. Expression of Cdt1 in G₂ induces replication in the presence of stable Cdc18. (A) FACS analysis of cdc25-22 cells transformed with pREP81-cdc18P (left) and pREP4-cdt1+ (right), shifted to 37°C in the absence of thiamine. (B) Dot plots of cells shown in (A), with forward scatter plotted against DNA content. (C) 2D gels of cells shown in (A) collected at 0 and 8 h after the shift to 37°C and probed for ars3001. (D) Boiled extracts from cells in (A) were western blotted for Cdc18. Tubulin serves as a loading control. (E) Boiled extracts were prepared from cdc25-22 cells transformed with pREP81-cdc18wt (left) and pREP4-cdt1+ (right), shifted to 37°C in the absence of thiamine. Samples were run on SDS–PAGE and western blotted for Cdc18. Tubulin serves as a loading control.