Figure 3.

TLR7 activation by dG does not require RNA. (A, B) BMDMs were treated with 0.5 mM dG for 24 h and supernatants were analysed as in Fig. 1A and B. Before the addition of dG, media were replaced where indicated with R10 (RPMI containing 10% FCS), R0 (RPMI with no additions), or fresh R9 (RPMI containing 10% FCS and 20% L929 conditioned medium). (C, D) BMDMs were differentiated from bone marrow in R9 (L929) or in R10 with 20 ng/mL M‐CSF. BMDMs were then treated with 0.5 mM dG, 2.5 µg/mL R837, 2.5 µg/mL CpG‐B DNA, or 2.5 ng/mL LPS for 24 h. Supernatants were analysed as in Fig. 1A and B. (E, F) dG or the indicated TLR agonists were pre‐incubated with (+) or without (−) benzonase (250 U/mL) for 30 min and were then added to BMDMs (0.5 mM dG, 2.5 µg/mL R837, 2.5 ng/mL LPS, 200 ng/mL poly I:C). Supernatants were analysed after 24 h as in Fig. 1A and B. Pooled data from biological replicates (BMDM cultures originating from individual mice, n = 3) are shown with mean ± SD. Data are representative of three (A, C) or two (B, D–F) independent experiments. Dashed lines represent detection limits. p‐values determined with two‐way ANOVA are indicated. ns, p $\ge$0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.