Nbs1 nb100 143

Immunoblots were performed as previously described (24 (link)). For immunoprecipitations, cells were washed with PBS and lysed on ice in TNG buffer (50mM Tris (pH 7.5), 200mM NaCl, 50mM NaF, protease inhibitors) for 30 min. Samples were pre-cleared with Protein A / Protein G agarose beads (Calbiochem, La Jolla, CA) and incubated with indicated antibodies or normal rabbit IgG and Protein A / Protein G agarose beads overnight, washed with TGN buffer and analyzed by immunoblot. Antibodies used were Nbs1 (NB100-143, Novus, Littleton, CO), Brca1 (OP92 and OP93, Calbiochem, La Jolla, CA).

Antibodies used for immunoblotting were: beta-actin (C4, Chemicon), ATM (MAT3; gift from Y. Shiloh, Tel-Aviv, Israel), pKAP1 S824 (Bethyl laboratories), pCHK2 T68 (Cell Signalling), pATM S1981 (Epitomics), Brca1 (OP92 and OP93, Calbiochem, La Jolla, CA), Nbs1 (NB100-143, Novus, Littleton, CO), Rap80 (70822, Abcam, Cambridge, MA), pBrca1 S1387 (A300-007A, Bethyl, Montgomery, TX), pBrca1 S1423 (2838, Abcam, Abcam, Cambridge, MA).

The following antibodies were used: Histone H3 (ab1791, Abcam); H3K4me1 (07-436, Millipore); H3K4me2 (07-030, Millipore); H3K4me3 (ab8580, Abcam); H3K9me1 (ab9045, Abcam); H3K9me2 (ab1220, Abcam); H3K9me3 (ab8898, Abcam); SUV39H1#1 (07-958, Millipore); SUV39H1#2 (Active Motif); MRE11 (NB100-142, Novus Biologicals); NBS1 (NB100-143, Novus Biologicals); ATM (Ab-3, Calbiochem); ATR (N19, SantaCruz); phospho-γH2AX (05-636, Millipore), UPF1 (generous gift of Dr. Claus Azzalin)^{62 (link)}; α - phospho-(Ser/Thr) ATM/ATR substrate (6966, Cell Signaling).