Composition and immuno-stimulatory properties of extracellular DNA from mouse gut flora

Figure 1 Mucus layer and eDNA in the mouse small intestine and colon. The first two columns are images obtained by optical microscopy with AB-PAS staining, and the third one are images obtained by laser scanning confocal microscopy with TOTO-1 staining. Pictures from the first two rows are the colon mucus layer, while those of the third row are the small intestinal mucus layer. A-F: Colon; G-I: Small intestine; A, D, G: AB-PAS 10×; B, E, H: AB-PAS 40×; C, F, I: TOTO-1 10 ×.

Figure 2 Principal component analysis plots for terminal restriction fragment length polymorphism profiles (including TRF size and relative abundance data). A: with AluI. B: with DdeI; Empty circle, eDNA; Empty square, iDNA.

Figure 3 Community structure component diagram at the phylum (A) and genus (B) levels. Samples were from eDNA (E) and iDNA (I) in triplicate, respectively. eDNA: Extracellular bacterial DNA; iDNA: Intracellular bacterial DNA.

Figure 4 Effect of iDNA and eDNA on cytokine production by Raw267. 4 cells, including TNF-α (A), IL6 (B), IL-12 (C), IL-10 (D), and ratio of TNF-α (E), IL6 (F) and IL-12 (G) to IL-10. Cells were treated for 12 h with medium, lipopolysaccharide (LPS) (1 μg/mL), eDNA (1 ng/mL), iDNA (1 ng/mL), eDNA(1 ng/mL) + LPS (1 μg/mL), or iDNA (1 ng/mL) + LPS (1 μg/mL) for 12 h. Values are expressed as mean ± SEM (n = 6). ^aP < 0.05, Significant differences between control and treatment; ^bP < 0.01, significant differences between control and treatment; ^cP < 0.05, significant differences between LPS treatment and treatment. E: Extracellular bacterial DNA; I: Intracellular bacterial DNA; LPS: Lipopolysaccharide.