Fig. 9: CpG DNA released during eructophagy activates vicinal cells. | Nature Communications

Fig. 9: CpG DNA released during eructophagy activates vicinal cells.

From: Macrophages disseminate pathogen associated molecular patterns through the direct extracellular release of the soluble content of their phagolysosomes

Fig. 9: CpG DNA released during eructophagy activates vicinal cells.The alternative text for this image may have been generated using AI.

a Schematic representation of experiment. DNA-conjugated experimental particles are phagocytosed, and particle-restricted DNA must be cleaved by lysosomal DNases prior to eructophagy-mediated extracellular release in order to activate vicinal HEK Blue TLR9 reporter cells through the TLR9-driven expression of SEAP (see text for details). b, d Detection of CpG-containing DNA fragments released into the supernatant by b PI3K−/− or WT, or d Beclin 1 Cre+/− or Beclin 1 Cre−/− BMMØs 4 h after phagocytosis of experimental particles covalently tethered to CpG-containing, 5.1 kb, plasmid DNA or BSA alone. Data are presented as copy number of plasmid DNA as detected by qPCR using a standard curve of CpG-containing, 5.1 kb, plasmid DNA at known concentrations. c, e Relative SEAP expression in HEK Blue Null cells or HEK Blue TLR9 cells co-cultured with either, c PI3K−/− or WT or e Beclin 1 Cre+/− or Beclin 1 Cre−/− BMMØs containing experimental particles covalently tethered to CpG-containing, 5.1 kb, plasmid DNA or BSA alone. Data are presented relative to the negative control HEK Null cells co-cultured with WT BMMØs containing plasmid DNA-tethered-experimental particles. b n = 7. c n = 3 or n = 4. d n = 3. e n = 3. Error bars presented as means ± SEM. *P ≤ 0.05 by one-way ANOVA with Bonferroni comparison of means. Source data are provided as a Source Data file.

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