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. 2015 Oct 2;99(3):455–465. doi: 10.1189/jlb.3A0515-201R

Figure 4.

Figure 4

R848 alone does not induce DNA strand breaks, but R848 and CTP synergistically enhance strand breaks and TNF‐α overproduction in FANCC‐deficient mononuclear phagocytes. (A) T‐shNT and T‐shFC cells were plated at a concentration of 105 cells/ml and treated with R848 (30 μM) or CPT (10 µM) for 2 h. (B) T‐shNT and T‐shFC cells were plated at a concentration of 105 cells/ml and pretreated with CPT (10 µM) for 2 h before stimulation with R848 (30 μM) for 2 h. Five thousand cells from each condition were analyzed using the comet assay. The mean ± sem percentage of tail DNA is shown. (C) T‐shNT and T‐shFC cells were plated at a concentration of 106 cells/ml and pretreated with CPT (100 nM or 1 µM) or MMC (100 mM, 300 mM, or 1 µM) before stimulation with R848 (30 μM) for 24 h. Secreted TNF‐α was measured in the conditioned media by ELISA. Data shown (mean ± sd) were derived from 3 biologic replicates for each condition, and P values were calculated using the paired 2‐tailed Student t test. CPT enhanced TNF‐α production only in R848‐stimulated T‐shFC cells but MMC had no such effect.