Levels of the ATM target proteins ph-p53 and γH2AX increase in association with increased global translation following XR. (A) Densitometry. Western bands (from Figure 6A) were quantitated and the raw data plotted. Red columns are baseline ph-p53 (following XR but without CD40L/IL-4); blue columns are levels of ph-p53 following XR and with CD40L/IL-4 stimulation. Patient ID is along the x-axis. There is a significant difference (paired 2-tailed Student t test) between ph-p53 levels at baseline and after stimulation (P < .0001). (B) Fold increase in ph-p53 after CD40L/IL-4 stimulation of X-irradiated cells correlated with fold-increase in viability due to CD40L/IL-4 (R² = 0.4, P = .03). (C) Cell viability (ATP luminescence RLU) following different doses of XR (5, 20, 35, and 50 Gy) in samples that were either untreated (orange lines) or CD40L/IL-4–treated (blue lines) (n = 10). Although there was no significant difference (paired 2-tailed Student t tests) in ATP luminescence between CD40L/IL-4–stimulated and unstimulated at 5 Gy (P = .06), there were significant differences at 20 Gy (P < .0001), 35 Gy (P < .0001), and 50 Gy (P = .0002). (D) Relative improvement in cell survival due to CD40L/IL-4 following XR. Cells were stimulated with CD40L/IL-4 and either treated with XR (50 Gy) or not treated. Viability (percentage as compared with no XR) correlated with fold-increase in viability due to CD40L/IL-4 alone (R² = 0.46, P = .03). The blue dot is a 17p deleted case (patient 46) demonstrating that this had among the highest viability following XR. (E) γH2AX response (percentage as compared with T = 0) in CLL cells at times up to 7 hours following XR (5 Gy) either stimulated with CD40L/IL-4 (blue lines) or unstimulated (orange lines) (n = 11). There were significant differences (paired 2-tailed Student t tests) in ATP luminescence between CD40L/IL-4–stimulated and unstimulated at 40 minutes (P = .0002) and 120 minutes (P = .0002), but not at 420 minutes (P = .06) (F) γH2AX response (percentage as compared with no XR) following doses of XR (2, 5, 10, and 20 Gy) in CLL cells either stimulated with CD40L/IL-4 (blue lines) or unstimulated (orange lines) (n = 5). There were significant differences (paired 2-tailed Student t tests) in γH2AX response between CD40L/IL-4–stimulated and unstimulated at 5 Gy (P = .01) and 10 Gy (P = .01). (G) CD40L/IL-4 response correlates with γH2AX response. The ratio of ATP luminescence in XR-treated and CD40L/IL-4–stimulated cells to that obtained in unstimulated but XR-treated cells is correlated to γH2AX response (determined as ratio of mean fluorescence index from XR-treated and CD40L/IL-4–stimulated cells to that from unstimulated but XR-treated cells) (n = 10) (R² = 0.59, P = .006). (H) OPP response correlates with γH2AX response (n = 10) (R² = 0.5, P = .015). The ratio of mean fluorescence index obtained in XR-treated and CD40L/IL-4–stimulated cells to that obtained in unstimulated but XR-treated cells is plotted for both OPP and γH2AX.