Figure 5.
Translational efficiency of ATM, MRN complex, and RIF1. (A) Mean translational efficiency for each gene (ATM, MRE11, RAD50, NBN, and RIF1) in type A or type B is presented as a column and the error bars represent SEM. All 5 genes have significantly different TE (Xtail analysis) between types A and B: ATM, P = .0117; MRE11, P = .0084; NBN, P = .0002; RAD50, P = .0038; and RIF1, P = .0009. (B) Western blots showing the effects of CD40L/IL-4 culture on expression of ATM, MRE11, NBN, RAD50, and RIF1 (n = 11). GAPDH is the loading control. Patient ID is given above (labeled 37-47) the autorads. (C) Densitometry. Western bands were quantitated and the raw data plotted. Red columns are baseline ATM; blue columns are levels of ATM following CD40L/IL-4 stimulation. Patient ID is along the x-axis. There is a significant difference (paired 2-tailed Student t test) between ATM levels at baseline and after stimulation (P = .001). (D) Comet assay. Unstimulated cells (solid red circles) and CD40L/IL-4–activated cells (white circles) were exposed to XR (5 Gy). Olive tail moment was measured at 0, 10, and 40 minutes. Horizontal bars are medians and error bars are interquartile ranges. Olive tail moment is significantly lower in CD40L/IL-4–activated cells (Mann-Whitney U test, ****P < .0001). Patient ID40. (E) Comet assays were carried out in a further 5 patients (IDs 41, 44, 45, 46, and 47) with or without CD40L/IL-4 stimulation. Olive tail moments at 40 minutes have been median normalized to facilitate comparisons. Horizontal bars are medians and error bars are interquartile ranges. There are significant differences in olive tail moment following activation by CD40L/IL-4 for patients ID41, ID44 and ID47, but not for patients ID45 and ID46. (Mann-Whitney U test; patient ID41, *P = .02; ID44, *P = .04; ID45, P = .07; ID46, P = .06; and ID47, ****P = .0001). ns, not significant.