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. 2011 Oct 10;10(10):1000-1; author reply 1002.
doi: 10.1016/j.dnarep.2011.07.013. Epub 2011 Aug 23.

CGK733 does not inhibit ATM or ATR kinase activity in H460 human lung cancer cells

Serah Choi, Luis I Toledo, Oscar Fernandez-Capetillo, Christopher J Bakkenist

CGK733 does not inhibit ATM or ATR kinase activity in H460 human lung cancer cells

Serah Choi et al. DNA Repair (Amst). .
No abstract available

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Figures

Fig. 1
Fig. 1
(A) Immunoblots of IR-inducible ATM kinase-dependent phosphorylations on ATM serine 1981 and CHK2 threonine 68. H460 human lung cancer cells were treated with 10 μM CGK733 (Tocris Bioscience), 10 μM KU55933 or 1 μM KU60019 (KuDOS Pharmaceuticals) from 30 min prior to exposure to 5 Gy γ-rays until harvest at 60 min following the insult. Briefly, whole cell extracts were prepared in lysis buffer: 50 mM Tris–HCl pH 7.5, 150 mM NaCl, 50 mM NaF, 1% Tween-20, 0.5% NP40 and 1× protease inhibitor mixture (Roche Applied Science, Indianapolis, IN). Cleared cell extracts were resolved in 3–8% Tris–acetate gels (Invitrogen, Carlsbad, CA) and immunoblotted with rabbit monoclonal anti-ATM 1981S-P antisera (EP1890Y, Epitomics), generic mouse monoclonal anti-ATM antisera (MAT3-4G10/8, Sigma), rabbit polyclonal anti-CHK2 68T-P (2661, Cell Signaling) and mouse monoclonal anti-CHK2 (1C12, Cell Signaling). (B) Immunoblots of the phosphorylation of ATM on serine 1981 and ultraviolet radiation (UV)-induced phosphorylation on CHK1 serine 317. H460 human lung cancer cells were treated with either 10 μM CGK733 or 10 μM ETP-46464 from 30 min prior to exposure to 10 J/m2 UV radiation until harvest at 6 h following the insult. Cleared cell lysates were immunoblotted with rabbit polyclonal anti-CHK1 317S-P (2344, Cell Signaling) and mouse monoclonal anti-CHK1 (2G1D5, Cell Signaling).
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