Atm 2873

Pools of small interfering RNA (siRNA) oligonucleotides directed against Rad54 (sc-36363), DNA-PKcs (sc-35201), or STAT3 (sc-29493) and control RNA (sc-37007) were purchased from Santa Cruz Biotech Inc. (Santa Cruz, CA, USA). Human (Hsa)-miR-21–5p (same sequence as mouse miR-21–5p) inhibitor was purchased from Ambion Inc. (Austin, TX). WT MEFs (control for Rad54 deficient cells (shown in Fig. 2a) and an additional WT line obtained from Dr Gloria Li’s lab (shown in Supplementary Fig. 2)) were transfected with oligonucleotides using Lipofectamine 3000 (Invitrogen, San Diego, CA, USA) according to the manufacturer’s instructions. Cells were then collected at 24–48 h after transfection and whole cell lysates were prepared in RIPA buffer as described previously [3 (link)]. Protein levels were determined using antibodies (against Rad54 (D-18), DNA-PKcs (H-163), EGFR (sc-03), STAT3 (sc-482) or actin (sc-47778)) that were purchased from Santa Cruz Biotech Inc. Antibodies against AP-1 (c-Jun) (#9165), phosphorylated EGFR (#3777) and ATM (#2873) were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). A phosphorylated ATR antibody (ABE462) was purchased from Millipore-Sigma Inc. (Billerica, MA, USA). The ATM inhibitor (KU-55933 (S1092)), ATR inhibitor (VE821 (S8007)), and EGFR inhibitor (gefitinib (ZD1839)) were purchased from Selleckchem (Houston, TX, USA).