Fig. 7: The regulatory effect of ATM on CBP and DOT1L is DNA damage independent.

MOLM-13 (A) and MV4-11 (B) cells were treated with hydroxyurea (Hu, 0.5 mM or 2 mM) for the indicated times. The lysates were extracted and detected by western blotting with the indicated antibodies. C The purified HA-CBP was subjected to in vitro phosphorylation with or without Flag-ATM(S1981A), and the reactions were then detected by western blotting with the indicated antibodies. D, E HA-CBP was cotransfected with FLAG-Vec, FLAG-ATM (WT), or FLAG-ATM(S1981A) into 293T cells. After 24 h, the cells were treated with or without 25 μM Ku55933 for 48 h. The cells were collected, and the cell lysates were then subjected to IP with HA antibodies. The input and IP samples were then detected by western blotting with the indicated antibodies (D). For the input samples, the band intensity of HA was normalised to that of ACTB in the same sample, and that of p-S/TQG was normalised to that of HA for the IP samples. The data from three independent experiments were used for statistical analysis (E). Student’s t test was performed for significant differences. ns not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. F. HA-DOT1L was cotransfected with or without HA-CBP into 293T cells. After 24 h, the cells were treated with or without 25 μM Ku55933 for 48 h. The cells were collected, and the cell lysates were then subjected to western blotting with the indicated antibodies. G. HA-CBP was cotransfected with or without FLAG-ATM into 293T cells. After 24 h, the cells were treated with or without 25 μM Ku55933 for 48 h. The cells were collected, and the cell lysates were then subjected to western blotting with the indicated antibodies. MOLM-13 (H) and MV4-11 (I) cells were treated with 25 μM Ku55933 for 20 h and 25 μM MG132 for 4 h. The cell lysates were then subjected to IP with CBP or DOT1L antibodies. The input and IP samples were then detected by western blotting with the indicated antibodies.