FIGURE 4.

SPDYA, CDK2, and Cyclin E are not associated with TTM complex. The expression plasmids of GFP, GFP-TERB1, GFP-TERB2, and GFP-MAJIN were co-transfected with FLAG-SPDYA (A), FLAG-CDK2 (B), FLAG-Cyclin E1 (C), or FLAG-Cyclin E2 (D) into HEK293T cells. Cells were harvested and lysed, 10% cell lysates were used as input, and the remained lysates were incubated with GFP-Trap. After incubation, the GFP-Trap was washed to remove unbound proteins. Then samples of input and Co-IP were subjected to western blotting.