Figure EV5. A mutant of E2F1 lacking a CY motif is not degraded by cyclin F at the G2/M transition (related to Fig 6).

1. HEK293T cells were cotransfected with MYC‐tagged ubiquitin and HA‐E2F1 or HA‐E2F1 ΔRxL lacking the CY motif as indicated (+). Indicated proteins were resolved by SDS–PAGE and detected by WB. H3 was used as a loading control. Correspond to the input of Fig 6B.
2. HeLa cells stably expressing HA‐E2F1 WT and HA‐E2F1 ΔRxL under the control of a retroviral promoter were synchronised by double thymidine block and harvested at the indicated hours (h) after release. Indicated proteins were resolved by SDS–PAGE and detected by WB. GAPDH was used as a loading control.