CC-115 inhibits the DNA damage repair pathway and TORK in CLL cells. (A-B) Freshly isolated CLL cells were incubated with the indicated concentration CC-115 for 30 minutes and irradiated (5 Gy), and lysates were made after 30 minutes. Protein lysates were probed for pDNA-PK (S2056), DNA-PK, pATM (S1981), ATM, pHSP90α (T5/7), HSP90α, yH2AX, and vinculin and cofilin for loading control. (A) Blots from 2 representative CLL samples are shown of 5 analyzed (supplemental Table 1; CLL patients 1, 2A, 3A, 4A, and 5). (B) Densitometric analysis of pDNA-PK, pATM, pHSP90α, and yH2AX is shown. ***P < .001 (1-way ANOVA). (C) CLL cells of ATM mutated patients (n = 4, patients 12-15) were treated with or without 1 µM NU7441 or CC-115 followed by irradiation (5 Gy), and yH2AX expression was measured at 2 hours using flow cytometry. *P < .05, **P < .01 (1-way ANOVA). (D) CLL cells were incubated with the indicated concentration of CC-115 for 30 minutes, and lysates were made after 30 minutes. Protein lysates were probed for pS6 (S240/244) and S6. Blot from patient 85 and densitometric analysis are shown (n = 3, patients 85, 87, and 88). Bars represent the mean ± standard error of the mean (SEM), ***P < .001 (1way ANOVA). (E) CLL cells were cultured in the presence or absence of 1 µM CC-115, CC-214, CC-292, or idelalisib for 1 hour. Protein lysates were probed for pS6 (S240/244) and actin for loading control. Blots from 3 representative CLL samples are shown (supplemental Table 1; CLL patients 19-21). (F) CLL cells (patient 20) pretreated with 1 µM idelalisib, CC-115, CC-214, and CC-292 were stimulated with αIgM for 20 minutes. Protein lysates were probed for pAKT(S473), pS6 (S240/244), p4EBP1 (T37/46), and pERK (T202/204) and actin as loading control.