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. 2012 Feb 8;40(10):4483–4495. doi: 10.1093/nar/gks041

Figure 2.

Figure 2.

Western blotting of DDR signaling kinases, partners and substrates. (A–D) MCF7 Cells were transfected with CUX1-specific siRNA. Nuclear extract were prepared and analyzed by immunoblotting. (A) Cells were exposed to 10 Gy of IR, incubated for 1 h prior to harvest and the extracts were immunoblotted for CUX1 to assess knockdown. (B) Cells were treated as in (A) and immunoblotted for ATM, p-ATM, 53BP1, Chk2 and p-Chk2. Actin was used to control for equal loading. (C) Cells were exposed to 20 Js of UV, incubated for 2 h prior to harvest and the extracts were immunoblotted for ATR, Chk1 and p-Chk1. (D) Cells were exposed to 10 Gy of IR, incubated for 6 h prior to harvest and the extracts were immunoblotted for p53 and CUX1. Actin was used to control for equal loading. (E) Nuclear extracts from Cux1Z/Z and wild-type MEFs were immunoblotted for ATM or ATR following exposure to 10 Gy of IR (left panel) or 20 Js of UV (right panel), respectively.