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. 2006 Jan 1;20(1):34–46. doi: 10.1101/gad.1381306

Figure 3.

Figure 3.

PIKK and CHK2 kinases are required for damage-induced BARD1–BRCA1 protein-binding events. (A) ATM-null lymphoblasts (–/–) (GM 03189D) and ATM wild-type lymphoblasts (+/+) (GM 03323A) were treated with DMSO (control) or the DNA-PK inhibitor, LY294002 (100 μM), as indicated. Endogenous BRCA1 was immunoprecipitated, and immunoblots were performed as noted. (B) DNA-PK-deficient glioblastoma cells (M059J) were treated with DMSO, or the indicated doses of caffeine and wortmannin and immunoprecipitated 1 h after 10 Gy. Immunoblots for BRCA1, phosphorylated BRCA1 at Ser 1423, and TopBP1 were performed as indicated. (C) HCT115 (CHK2-deficient) cells, expressing eBARD1, were reconstituted with either wild-type HA-tagged CHK2 or a vector control and were immunoprecipitated with Flag Ab 2 h after exposure to 10 Gy. Immunoblotting was subsequently performed as indicated.