Fig. 5: ATM confers CBP stability by preventing its ubiquitination-mediated degradation.

The total lysates of control (shSCR) and ATM knockdown (shATM) MOLM-13 (A) and MV4-11 (B) cells were subjected to western blotting with the indicated antibodies. C MOLM-13 and MV4-11 cells were incubated with 40 μg/ml CHX for the indicated times. The total lysates of the cells were then subjected to western blotting with the indicated antibodies. D MOLM-13 and MV4-11 cells were incubated with DMSO (vehicle, Veh) or 50 μM chloroquine (CHQ) for 24 h. The total lysates of the cells were then subjected to western blotting with the indicated antibodies. E MOLM-13 and MV4-11 cells were incubated with 40 μg/ml CHX in combination with DMSO (vehicle, Veh) or 25 μM MG132 for 6 h. The total lysates of the cells were then subjected to western blotting with the indicated antibodies. Control (shSCR) and ATM knockdown (shATM) MOLM-13 (F) and MV4-11 (G) cells were treated with 25 μM MG132 for 4 h. The cell lysates were subjected to IP with CBP antibodies. The input and IP samples were then detected by western blotting with the indicated antibodies. H 293T cells were cotransfected with HA-CBP and control vector (Flag-Vec) or Flag-ATM. At 36 h posttransfection, the cells were treated with 25 μM MG132 for 20 h. The cell lysates were subjected to immunoprecipitation (IP) with HA antibodies. The input and IP samples were then detected by western blotting with the indicated antibodies. I, J Flag-CBP was transfected into control (shSCR) or ATM knockdown (shATM) 293T cells. After 72 h, the cells were collected, and the cell lysates were subjected to western blotting with the indicated antibodies (I). The band intensity of ATM/DOT1L/CBP was normalised to that of ACTB in the same sample, and the data from three independent experiments were used for statistical analysis (J). Student’s t test was performed for significant differences. ns not significant. **P < 0.01, ***P < 0.001, ****P < 0.0001. K, L HA-DOT1L with or without Flag-ATM were cotransfected into control (shSCR) or CBP knockdown (shCBP) 293T cells. After 72 h, the cells were collected, and the cell lysates were subjected to western blotting with the indicated antibodies (K). The band intensity of ATM/HA-DOT1L/CBP was normalised to that of ACTB in the same sample, and the data from three independent experiments were used for statistical analysis (L). Student’s t test was performed for significant differences. ns not significant. **P < 0.01, ***P < 0.001, ****P < 0.0001.