Figure 6. Cdh1-promoted K11-linkage-specific ubiquitination of E2F1 is enhanced and accumulated after DNA damage. HEK293T cells were transfected with Flag-E2F1, HA-Ub-K11 and GFP-Cdh1 (or GFP vector control) as indicated. The cells were either left untreated or treated with 5 μM adriamycin for 5 h before harvesting. The lysates were prepared and analyzed for E2F1 ubiquitination as described in methods section. Fractions of the total lysates were analyzed as well (lower panels).