Figure 8

Independent non-overlapping regions are sufficient for Mcm10 proteolysis during M-phase. U2OS cells expressing different regions of Mcm10 were arrested in M-phase using nocodazole (0 h) and then harvested at the indicated time-points after release into drug-free medium. The proteolysis of different regions of Mcm10 evaluated after release from nocodazole by immunoblotting with an anti-HA antibody has been shown in parts (A), (C) and (E) and the flow cytometry of propidium iodide-stained DNA of stable cell-line expressing the corresponding fragment has been shown in parts (B), (D) and (F) respectively. ASN refers to samples from asynchronously growing cells while NS points to a non-specific band that displays equal protein loading. The observed mobility of fragments of Mcm10 (fused to GFP and HA epitope tags) was as follows: 607-707: 52 kDa; 770-875: 52 kDa; 770-783: 42 kDa; 843-875: 42 kDa; 783-823: 44 kDa; 803-843: 44 kDa; 783-803: 40 kDa; 803-823: 44 kDa; 823-843: 43 kDa.