Figures & data
Figure 1. Determine the kinetics of phosphorylation of the DNA damage-associated proteins treated with NCS. BJ cells (A), HCT116 cells (B), GM09607 cells (C) and HCT116 Chk2(-) cells (D) were treated with NCS (1 μg/ml) for the indicated time (from 1 min to 6 hours). Cell lysates were collected and protein phosphorylation was studied using the antibodies indicated. Actin serves as a loading control.
Figure 2. Determine the kinetics of phosphorylation of the DNA damage-associated proteins treated with Dox. HeLa cells were treated with NCS (1 μg/ml) or Dox (8.5 μM) for the indicated time (1 min to 6 hours). Cell lysates were collected and protein phosphorylation was studied using the antibodies indicated. Dox does not induce phosphorylation of Chk2 S33/35. Actin serves as a loading control.
Figure 3. Phosphorylation of the specific residues of Chk2 regulates other phosphorylation sites dependent on the DNA damage. HCT116 Chk2(-) cells were transduced by retrovirus expressing Chk2 cDNA of HA-tagged wild type, S19A, S33/35A or T68A. Stable clones were treated with NCS (A), 1 μg/ml) or Dox (B), 8.5 μM) for the indicated time. Cell lysates were collected and subjected to immunoblot analysis of the indicated antibodies. Actin serves as a loading control.
