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. 2001 Jan 2;98(2):497–501. doi: 10.1073/pnas.240461397

Figure 1.

Figure 1

Cleavage of cyclin A by cyclin B translated in rabbit RL. (A) Cyclin B(RL) induces efficient cleavage of cyclin A(CΔ114). GST-cyclin A(CΔ114) was mixed with unprogrammed RL (lanes 2 and 3) or cyclin B(RL) (lane 4). The reactions were incubated at 37°C and stopped with SDS-sample buffer at the indicated time. The proteins were analyzed by SDS/PAGE and Coomassie blue staining. Molecular size standards (lane 1) in kDa are indicated on the left. The positions of GST-cyclin A(CΔ114) and the cleaved form of cyclin A (*) are indicated on the right. (B) Cleavage of cyclin A expressed in mammalian cells into two fragments by cyclin B(RL). FLAG-cyclin A was transiently transfected into HtTA1 cells. Cell extracts were prepared and the expressed cyclin A were immunoprecipitated with anti-FLAG antibodies. The immunoprecipitates were incubated with unprogrammed RL or cyclin B(RL) as indicated. The samples were applied onto 12.5% SDS/PAGE (top two panels) or Tricine gel (bottom two panels), and subjected to immunoblotting with anti-FLAG mAb M2, anti-cyclin A monoclonal E23, or M2 and E23 together as indicated. In this paper, the N-terminal fragment of cyclin A is denoted with “*”, and the C-terminal fragment is denoted with “**” (see main text).